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Pivot
point
C H A P T E R 6 2 Pelvic Fixation of the Aging Spine
Pivot point
415
A
F IG UR E 6 2- 3   Sagittal (A) and axial (B) cross-sectional cuts through the pelvis depicting the location of the lum-
bosacral pivot point (red dot) along the posterior aspect of the L5-S1 interspace.
B

BASIC SCIENCE AND BIOMECHANICAL STUDIES

A thorough understanding of the osseous and soft tissue anatomy of the lumbosacral spine and pelvis is critical for the safe and effective inser­tion of iliac fixation. The sacrum is composed of five fused vertebrae and serves as a functional keystone that links two paired hemipelves posteri­orly through bilateral sacroiliac articulations. Because this instrumentation is introduced into the iliac wings, which form the lateral borders of the pelvic ring, it is necessary to consider a number of different parameters such as intrailiac distance, width between the inner and outer tables, and cortical thickness. The longest intrailiac path proceeds along a line con­necting the posterior superior iliac spine to the anterior inferior iliac spine, which averages approximately 141 mm in males and 129 mm in females. Similarly, the width of the iliac crest should allow for the placement of 8-mm screws in males and 6- to 7-mm implants in females; the mean cor­tical thickness in males and females has been shown to be 5.2 mm and
4.7 mm, respectively. In their comparison of 10 different methods of lumbosacral fixation,
McCord and colleagues the highest loads to failure both obtained solid purchase within the iliac crests. Based on this in vitro data, the concept of the lumbosacral pivot point was elucidated, which is located at the intersection of the middle osteoligamentous column and the L5-S1 interspace (Figure 62-3). Accord­ing to this paradigm, pelvic implants that extend anterior to this fulcrum increase the overall stiffness of the instrumentation because the resultant biomechanical vectors are transformed from in-line forces to cantilever
6
bending.
Three distinct zones have been designated for the purpose of catego-
rizing sacropelvic fixation (Figure 62-4). bral body, Zone 2 encompasses the sacral alae and the region between S2 and the coccyx, and Zone 3 denotes the paired iliac wings. In general, the degree of stability imparted by these screws has been found to improve as they are positioned more laterally within the lumbopelvic ring (i.e., from Zone 1 to Zone 3); implants situated in Zone 3 are also better able to resist the pull-out forces that are generated by bending moments arising from the lumbosacral junction than those directed toward either Zone 1 or 2.
The use of internal fixation in the setting of osteoporosis may be partic-
ularly problematic because this instrumentation is likely to be more suscep­tible to loosening or failure, especially in cases involving traumatic injuries or deformity correction where it may be subjected to tremendous stresses and strains. Consequently, in these situations it may be preferable to employ longer implants that will traverse the entire length of the iliac column and engage the cortical bone superior to the acetabulum, so that they are able to withstand more significant forces. cated for osteoporotic patients is the incorporation of two screws into each iliac crest. However, there are some concerns that the application of rigid
3
5
reported that the two constructs associated with
7
Zone 1 refers to the S1 verte-
2
Another approach that has been advo-
Zone 3
F IG UR E 62 - 4  Schematic demarcating  the  three  zones  of  sacropelvic 
fixation: 1—S1  vertebral  body;  2—the  region between  S2  and  the  coccyx as  well as the sacral alae; 3—iliac wings.
iliac fixation could lead to stress shielding of an already compromised pelvis, which may predispose these individuals to insufficiency fractures.
Zone 1
Zone 2
Zone 3
8

CLINICAL PRACTICE GUIDELINES

Unfortunately there are currently no prospective, randomized, controlled clinical trials that have specifically elucidated the safety and efficacy of lumbopelvic fixation in the elderly population. Consequently, the precise indications for this surgical technique have not yet been definitively estab­lished, and there are still no validated clinical practice guidelines that may be employed to direct the management of these patients.

CLINICAL CASE EXAMPLES: TREATMENT AND FUTURE CHALLENGES

Both of the cases presented earlier involved posterior-based spinal con­structs that were reinforced with pelvic implants. The first patient with degenerative scoliosis underwent a circumferential procedure performed
416
P A R T V I I Surgical Treatment Modalities: Lumbar Spine
in a staged fashion consisting of anterior releases and interbody fusions from L2-S1 with polyetheretherketone (PEEK) cages followed by a poste­rior spinal arthrodesis extending from T5 to the ileum with instrumenta­tion and autogenous bone (Figure 62-5). In the second case example, the pathologic fractures of the L3 and L4 vertebral bodies were addressed with a posterior decompression of those levels, in conjunction with an instru-
mented fusion of the motion segments between L1 and the pelvis with local and iliac crest autograft (Figure 62-6). In both of these situations the decision was made to include supplemental pelvic fixation because such extensive constructs would be expected to subject the distal screws to substantial forces that may bring about hardware failure or loos­ening, especially in these individuals who were suspected of having
F IG UR E 6 2 -5   Postoperative  PA  (A)  and  lateral  (B)  x-rays  following  a 
 circumferential arthrodesis with extension of instrumentation into the pelvis.
A
B
A B
F IG UR E 6 2- 6  Postoperative AP (A) and lateral (B) x-rays demonstrating a posterior lumbar construct 
that was augmented with iliac screws for greater stability.
C H A P T E R 6 2 Pelvic Fixation of the Aging Spine
417
osteoporosis. Thus the purpose of adding pelvic implants was to aug­ment the rigidity of the instrumentation and create a more stable, “balanced” construct that would hopefully reduce the incidence of pseudarthrosis.
In placing pelvic screws, it may be useful to remove a portion of the
posterior superior iliac spine with a Leksell rongeur or burr so that the heads may be adequately countersunk in an attempt to minimize their prominence. The probe is introduced into the window of cancellous bone and aimed between the inner and outer tables of the ileum, toward the sciatic notch (Figure 62-7). After the tract is probed for any cortical breaches, the cavity is tapped in preparation for the instrumentation; in most instances the dimensions of the iliac wing should be able to accom­modate relatively large implants (i.e., 7.5 mm in diameter and up to 80 mm in length or greater). The final position of the pelvic screws should be assessed with intraoperative radiographs or fluoroscopy in AP, lateral,
and oblique views to ensure that it is entirely contained within the ileum and is not encroaching upon other critical structures, such as the hip joint (Figure 62-8).
In the future, further advances in technology may yield even stronger implants that are lower profile and easier to connect to adjoining lumbosa­cral instrumentation. Because of the challenges associated with the inser­tion of iliac screws, this approach may be well suited to the application of novel surgical navigation systems that are integrated with advanced three­dimensional imaging modalities.

CONCLUSION

Pelvic fixation represents an effective method for enhancing the rigidity of posterior spinal fusion constructs. By obtaining secure purchase within the iliac crests, these implants confer greater stability to the axial skeleton
F IG UR E 62 - 7  Lateral view of the pelvis delineating the  optimal  trajectory  for  the 
screw.
A B
F IG UR E 6 2- 8  Anteroposterior (A) and oblique (B) fluoroscopic images are used to visualize the iliac screws dur-
ing the  procedure  to  ensure that the implants have  not  breached  the cortical bone and  are  not  encroaching  upon the  acetabulum.
418
P A R T V I I Surgical Treatment Modalities: Lumbar Spine
and may therefore be indispensable for the treatment of patients with osteoporosis who are at greater risk for nonunion and other postoperative complications following complex spinal reconstructions.

References

1. A. Moshirfar, F.F. Rand, P.D. Sponseller, S.J. Parazin, A.J. Khanna, K.M. Kebaish, J.T. Stinson, L.H. Riley, Pelvic fixation in spine surgery. Historical overview, indications, biomechanical rel­evance, and current techniques, J. Bone Joint Surg. Am. 87 (2005) 89–106.
2. R .M. Schwend, R. Sluyters, J. Najdzionek, The pylon concept of pelvic anchorage for spinal
instrumentation in the human cadaver, Spine 28 (2003) 542–547.
3. T.A. Schildhauer, P. McCulloch, J.R. Chapman, F. A. Mann, Anatomic and radiographic con­siderations for placement of transiliac screws in lumbopelvic fixation, J. Spinal Disord. Tech. 15 (2002) 199–205.
4. N.K. Acharya, B. Bijukachhe, R.J. Kumar, V.K. Menon, Ilio-lumbar fixation—the Amrita technique, J. Spinal Disord. Tech. 21 (2008) 493–499.
5. D.H. McCord, B.W. Cunningham, Y. Shono, J.J. Myers, P.C. McAfee, Biomechanical analysis of lumbosacral fixation, Spine 17 (Suppl. 8) (1992) S235–S243.
6. E.R. Santos, M.K. Rosner, J.H. Perra, D.W. Polly, Spinopelvic fixation in deformity: a review, Neurosurg. Clin. N. Am. 18 (2007) 373–384.
7. M.F. O’Brien, Sacropelvic fixation in spinal deformity, in: R.L. DeWald (Ed.), Spinal deformities: the comprehensive text, Thieme, New York, 2003, pp. 601–614.
8. K.B. Wood, M.J. Schendel, J.W. Ogilvie, J. Braun, M.C. Major, J.R. Malcom, Effect of sacral and iliac instrumentation on strains in the pelvis: a biomechanical study, Spine 21 (1996) 1185–1191.
Intradiscal Biologics: A Potential Minimally Invasive Cure for Symptomatic Degenerative Disc Disease?
Rajeev K. Patel
63
k e y p o i n t s
Biological repair of injured tissues by introducing cell-based tissue
replacements, genetic modifications of resident cells, or a combination thereof have been successfully applied to various tissues such as bone and cartilage.
Application of these treatment modalities to cure symptomatic intervertebral
disc degeneration is in its infancy and mostly limited to experimental studies in vitro or in animal studies.
Attempts at gene therapy or tissue engineering demonstrate obvious
potentials as well as significant shortcomings to biological cure of symptomatic degenerative disc disease.
Knowledge gained from these attempts might be applied to cure the low back
pain that often accompanies the structurally compromised intervertebral disc.

INTRODUCTION

The development of a cell-based, biological replacement to restore, main­tain, and improve the function of damaged tissues and organs has become the en vogue frontier to developing potential novel approaches to patient cures. The intervertebral disc (IVD) undergoes very extensive degenerative changes (Figure 63-1) with the various macro- and microtraumas that come with age and daily life activities. Individual differences have been demon­strated in young individuals exhibiting the disc of an elderly person and vice versa. It is generally accepted that an extremely prevalent rate and degree of asymptomatic disc degeneration exists in the general population. There­fore, differentiating normal aging from symptomatic pathological degenera­tion is very difficult and cannot be assessed by simply identifying the most abnormal disc on imaging (Figure 63-2). At this time, controlled provocative lumbar discography (Figure 63-3) remains our best clinical test to identify a physiologically painful structurally compromised IVD. The term ‘‘disco­genic low back pain” is the term often used to indicate degenerative disc disease associated with concordant pain.
Relating recent findings regarding the molecular mechanisms in initiat­ing or propagating degenerative alterations of the IVD will be crucial to the ultimate success in the developments of biologic strategies to cure disco­genic low back pain.
disc axially. The annulus fibrosus is made up of several lamellae consisting of parallel collagen fibers interspersed by elastin fibers. Surrounded by the annulus fibrosus is the nucleus pulposus, the gelatinous core, which consists of randomly organized collagen fibers, radially arranged elastin fibers, and a highly hydrated aggrecan-containing gel. The highly hydrated proteoglycans in the nucleus pulposus are essential to maintain the osmotic pressure and therefore have a major effect on the load-bearing properties of the disc.
It is also important to note that the intervertebral disc is a largely avascular structure. With increasing age, as growth and skeletal matura­tion proceed, degenerative processes begin to change the morphology and therefore the function of the disc. The most widely accepted conceptual model of spinal segmental degeneration was proposed by Kirkaldy-Willis. In this model, the nucleus pulposus of degenerated discs is characterized by a decreased water and proteoglycan content leading to the loss of its gel-like appearance and hydrostatic properties. Degenerative changes of the annulus fibrosus are less obvious, but result in irregular lamellae with the collagen and elastin networks becoming more disorganized. Replacing the gel-like structure of the nucleus pulposus with fibrocartilaginous tissue results in decreased flexibility and therefore often in cleft formation with fissures. Up to 50% of the cells show signs of necrosis and some of them reveal signs of apoptosis, potentially resulting in cell loss from the disc. Although there is broad consensus about these hallmarks of degeneration, the question of whether revascularization and/or reinnervation of the inner parts of the disc may occur during degeneration is still a topic of debate. Although studies have described revascularization, possibly accompanied by reinnervation, of the inner parts of the IVD, it is not completely clear at
Normal disks Disk degeneration
1
2
3

FUNCTIONAL ANATOMY OF THE INTERVERTEBRAL Disc

Intervertebral discs transmit loads from body weight and muscle activity as well as provide flexibility to the spine. The discs consist of three highly specialized structures: the endplates, the annulus fibrosus, and the nucleus pulposus The two cartilaginous endplates form the inferior and superior interface between the disc and the adjacent vertebrae, thereby enclosing the
F IG UR E 63 -1   Macroscopic  pathoanatomy  evident  in  disc  degenera-
tion compared side by side to healthy intervertebral discs.
419
420
F IG UR E 6 3- 2   T2-weighted  sagittal  MRI  demonstrating  segmental 
degenerative disc desiccation and bulging at the L4-L5 level.
P A R T V I I     Surgical Treatment Modalities: Lumbar Spine

CAUSES OF DEGENERATIVE DISC DISEASE

Degenerative disc disease is a complex process with a multifactorial etiol­ogy. Nutritional effects, mechanical load, and genetics all likely have con­tributory pathologic effects on the IVD. Of these, nutrition and removal of waste products likely play a special role in realizing the potential that intradiscal biologics may ultimately hold. Insufficient nutritional supply of the cells is thought to be the major obstacle contributing to degenera­tive disc disease. Cells of the IVD face the precarious situation of having to maintain a huge extracellular matrix with a ‘’fragile’’ supply of nutrients that is easily disturbed because the IVD is avascular and nutrition is depen­dent on diffusion. Because of the size of the intervertebral disc, the nutrients need to diffuse from a capillary network in the vertebral bodies, through the endplates and the disc matrix to the cells in the nucleus of the disc. The supply of nutrition becomes more restricted as the originally carti­laginous endplates become calcified as the degenerative process progresses. As glucose and oxygen is restricted because of diffusion distances, the removal of metabolic waste such as lactic acid becomes critically impaired. Measurements have demonstrated that as oxygen concentrations were very low in the nucleus and increased toward the disc surface, the lactic acid con­centration showed the reverse profile. The buildup of lactic acid results in an intradiscal environment with a lowered pH. Low oxygen concentrations and acidic pH adversely affect the synthetic activity and proteoglycan synthesis rates of disc cells. This toxic environment may lead to a fall in proteoglycan content and therefore to degenerative disc disease. This suboptimal environ­ment may lead to increased cell death and therefore reduced cell numbers in the disc. that very few remaining cells are confronted with the task of maintaining an extensive matrix. Unfortunately, it likely holds true that the progression of matrix degeneration becomes irreversible once the cell density falls below a minimal threshold.
4
Ultimately, the result of poor nutritional supply of the IVD is
L3–4: Normal nucleogram .7 cc. injected firm resistance 0/10 pain
L4–5: Normal nucleogram .6 cc. injected non-concordant pain 3/10 in back
L5–S1: DDD .9 cc. injected concordant pain back and R leg 8/10
F IG UR E 6 3- 3  Provocative lumbar diskography demonstrating a struc-
turally  compromised  fissured  L5-S1  IVD  with  a  physiologic  concordant  pain  elicitation  under  controlled  pressure  as  well  as  adjacent  nonphysiologic  and  nonpainful discs at L4-L5 and L3-L4 with intact morphologies.
which stage of degeneration these occur.3 Clarification of this question is of special importance since the interplay between neovascularization and neo­innervation might be of crucial importance for the pain sensation caused by degenerated discs. Answers to these questions may ultimately determine the rate-limiting step to the potential of biologic cures of symptomatic degenerative disc disease.

THERAPEUTIC BIOLOGIC STRATEGIES

Biologic treatments for the degenerated IVD have scarcely been utilized to regenerate or cure the painful, deteriorated disc and restore biological func­tion. Thus far, intradiscal biologics are classified into four approaches:
1. Direct injection of a biologically active factor(s) (Figure 63-4)
2. Modification of the gene expression of resident disc cells in vivo (direct gene therapy) (Figure 63-5)
3. Supplementation with autologous implantation of in vitro cultivated and modified cells (Figure 63-6)
4. Stem-cell based gene therapies (Figure 63-7)
These applications aim for sustained delivery of biologically active sub­stances to the disc that should drive regeneration or conserve the status quo. The nature of the respective active factor is hereby defined by our knowledge of the molecular mechanisms active in the disc during the various stages of degeneration. The applicability of the various approaches is largely depen­dent on our current knowledge of disc cell biology, the state of degeneration of the intervertebral disc, and potential safety issues.
Intradiscal Injection of a “Naked” Biologically Active Factor
Percutaneous intradiscal injection would provide the most straightforward approach to delivery of an active biologic factor to the disc cells (see Figure
63-3). Although direct application of potentially beneficial factors, mostly
proteins like growth factors, cytokines, or anabolic enzymes, has been used frequently in vitro, few studies have been published attempting this approach in vivo. Promising results have been reported after injecting rabbit lumbar IVD in vivo with osteogenic protein-1 (OP-1), a growth factor belonging to the Transforming Growth Factor Beta (TGF-b) superfamily of growth fac­tors. Direct injection of this growth factor resulted in significantly increased proteoglycan synthesis and restoration of disc height that was found to be stable up to 24 weeks after injection. that OP-1 injection exhibited a physiologic effect by inhibiting pain-related behavior in a rat disc degeneration model. colleagues
9
documented the anticatabolic effect of intradiscal injection of
5,6
Additional studies demonstrated
7,8
Subsequently, Chubinskaya and
C H A P T E R   6 3     Intradiscal Biologics: A Potential Minimally Invasive Cure for Symptomatic Degenerative Disc Disease?
Induction of
FI G U RE 63 - 4  Intradiscal injection of a “naked” biologically active factor—done to facilitate the sustained release 
of an agent into the cellular matrix.
FI G U RE 63 -5   Modification of the gene expression of resident disc cells in vivo (direct gene therapy) utilizing a viral 
vector resulting in transformation and sustained expression of the active protein Z.
Harvesting of
disk cells
Cultivated cells
Seeding of
scaffold
Transformed cells
421
Cell-seeded
scaffold
FI G U RE 63 -6   Supplementation with autologous implantation of in vitro cultivated and modified cells. Cultivated 
cells  can  be  genetically  modified  in  vivo  before  implantation  (indirect  gene  therapy),  seeded  into  a  scaffold,  or  simply  implanted directly.
Harvesting of
bone MSCs
FI G U RE 6 3- 7  Stem-cell based gene therapy.  Mesenchymal  stem  cells  can  be  cultivated  as  progenitor cells and 
either injected directly into the disc matrix or be differentiated in vitro into a disc cell and then injected.
Progenitor
OP-1 in a rat model by demonstrating reduced immunostaining for aggre­canase, Matrix Metalloproteinase (MMP)-13, substance P, Tumor necrosis factor (TNF)-α, and Interleukin (IL)-1β. Because substance P is a neuro­peptide linked with inflammation and pain, the aforementioned reduction in level of this noxious protein support the previously stated physiologic inhi­bition of pain-related behavior.
7-9
Furthermore, Miyamoto and coworkers10 were able to demonstrate that intradiscal injection of OP-1 restored the biomechanical properties of IVDs in the rabbit model of degenerative disc disease. They reported not only that a single injection of OP-1 significantly
Implantation of
autologous cells
differentiation
cells
Injection of
MSCs
Differentiated
cells
Injection of differentiated cells
restored IVD height, but also that the treated discs demonstrated a higher viscous and elastic modulus due to increased proteoglycan content in the nucleus as well as increased collagen content in the nucleus and annulus. Concerns regarding the potential of ectopic bone formation in the epidural space with OP-1 therapies was addressed by Kawakami and associates. They demonstrated that there was no macroscopic evidence of ectopic bone formation, no motor paresis, and no behavioral differences to motor stimuli with epidural administration of OP-1 in a rat model. The aforementioned studies demonstrate the feasibility of direct injection, yet this technique
11
422
P A R T V I I     Surgical Treatment Modalities: Lumbar Spine
may be limited to the presence of disc cells that are still healthy, numerous, and able to respond to a biologically active agent. Taking into context the decreasing viability and synthetic activity of human disc cells during pro­gressive degeneration, future directions for this technique may be best suited for success in the younger patient population with discogenic Low back pain (LBP) due to modestly degenerated discs in synergistic combinations with other biologically active agents.
As opposed to injecting a biologically active enzyme or growth factor,
Klein and colleagues
12
published a clinical pilot study utilizing direct injec­tion of a mixture of matrix components and aiding components known to induce proteoglycan synthesis. This solution of glucosamine and chondroi­tin sulfate combined with hypertonic dextrose and dimethylsulfoxide was injected into 30 patients who exhibited concordant pain on provocative lumbar discography. These patients responded well regarding reduction in a disability score and a visual analogue pain score at an average follow-up of 13 months. The authors suggested that the good outcome might be due to the combination of several components resulting in a parallel replenishing of the matrix by increased proteoglycan synthesis and induction of disc repair by simultaneous induction of multiple growth factors. This approach might prove superior to the injection of a single bioactive factor. It is conceivable that the injected matrix components are able to modulate and improve the intradiscal environment, enabling the disc cells, even in a degenerated disc, to react to the resulting secretion of growth factors and continue with the maintenance of the cellular environment. However, from this pilot study it is not clear if the injected components will be contained inside the disc in heavily degenerated discs and therefore be able to ensure a prolonged beneficial effect on the disc cells. Further controlled comparative studies are required before any therapeutic conclusions can be rendered.
Common to these injection techniques is the concern that the afore­mentioned demonstrated short-term effects might cease when the originally injected material has been consumed or is lost to the disc cells by diffusion. In order to provide the disc with a continuing supply of biologically active factors, it would be desirable to continuously produce the biologic of choice or include the substance in a pharmacological slow-release system as sug­gested for the use of growth factors.
13
Considering these data, it is conceiv­able that combination therapy of growth factor(s) and matrix replenishment might be the way to obtain a more sustained improvement of degenerated discs. This approach may also allow for expansion of the previously stated limitations with application of this technique to various grades of degenera­tion because of the requirement of a certain density of healthy disc cells.
Gene Therapy Approaches
Prolonged supply of the discal matrix with a beneficial agent could be achieved by genetically modifying disc cells to produce a desired gene prod­uct. Because of advances in molecular genetics, techniques are readily avail­able to insert genetic elements (DNA) into almost any type of target cell. These genetic elements usually consist of the gene encoding the desired product and a control element modulating the expression of the respective gene. Typically, two strategies can be used to achieve expression of the desired gene at the targeted site. Direct, or in vivo, gene therapy requires the direct introduction of the gene of interest into resident cells in situ (see Figure
63-4), whereas indirect, or ex vivo, gene therapy requires the removal of tar-
get cells, introduction of the gene of interest in vitro and implantation of the transformed cells (see Figure 63-5). usually in the form of pure “naked” DNA, can be optimized by application of a carrier, also called vector. Viral vectors are very efficient transporters of genetic material; they are able to enter mammalian cells, taking over DNA replication and the protein expression machinery. For the purposes of gene therapy, several engineered viruses are available that have the original viral genome removed or inactivated and, in addition, are modified to not replicate or exhibit their pathogenicity. Of interest, these viruses vary in their ability to integrate the transferred DNA into the host cell genome, their ability to invade dividing or nondividing cells, and their infection efficiency. Because of the properties of the IVD and its cells, the virus of choice needs to efficiently infect nondividing, quiescent cells. Furthermore, the low cell density inside the disc might hamper efficient infection of a sufficient fraction of the disc cells. On the other hand, the avascular and contained IVD might provide an advantageous environment to achieve high concentrations of the injected
14
Uptake of the desired genetic material,
viral vector, leading to higher efficiency of the infection process while also lessening the danger of an immune response against viral proteins.
Studies have demonstrated that adenoviral vectors are able to efficiently transform disc cells from various species. The main disadvantage of adeno­viral vectors lies in the activation of innate and adaptive parts of the patient’s immune system when the vector is applied in vivo. To overcome this poten­tially lethal complication, Lattermann and colleagues
15
recently tested an adeno-associated viral vector (AAV) for its applicability to degenerative disc disease. The authors demonstrated that AAV was able to efficiently transduce human disc cells in vitro and rabbit disc cells in vivo. Although AAV caused a humoral immune response, no significant cellular immune response, as seen with adenoviral vectors, was observed. Interestingly, despite the observed humoral immune response, significant transgene expression was observed in the preexposed animals.
15
These findings suggest that AAV might offer a valuable and safer alternative to adenoviral vectors in the future. Although the aforementioned studies suggest the feasibility of direct gene therapy using viral vectors to target disc cells, the question of the delivered gene and therefore expressed gene product remains open. Nishida and coworkers
14
published one of the initial studies to deliver an exogenous therapeutic gene in vivo using an adenoviral vector carrying the gene for TGF-b1 to modify cells of rabbit IVD. The authors found significant increases of TGF-b1 and proteoglycan produc­tion in the injected disc, suggesting the feasibility of direct gene therapy to treat intervertebral disc degeneration.
14
Lim Mineralization Protein (LMP) is another potentially beneficial gene factor that has been shown to positively affect the degenerated disc cellular matrix by increasing the disc cell produc­tion of Bone Morphogenetic Proteins (BMP) and proteoglycans in vitro.
16
In vivo studies involving the intradiscal injection of rabbit discs resulted in increased expression of the anabolic cytokines BMP-2 and BMP-7 mRNA and also led to increased production of aggrecan mRNA.
16
These data sug­gest that LMP-1 is also a beneficial factor that could be applied to gene thera­peutics for disc degeneration. Sox9-on the other hand, does not affect the proteoglycan content of the disc cellular matrix. Sox9 is a gene transcription factor responsible for the synthesis of type II collagen, and its transfer into cells from degenerated human discs resulted in increased levels of type II col-
17
lagen.
Injection of a Sox9-carrying adenoviral vector into traumatized rab­bit discs resulted in preservation of the histologic appearance seen in healthy discs, whereas the injured control discs displayed typically recognized degen­erative changes.
17
Therefore, not only increased proteoglycan production but also collagen type II production seems to be able to prevent disc degeneration in vivo, thereby offering multiple potentially therapeutic options. Although the increased production of a single gene product seems to result in the trans­formation of disc cells, a combination of related gene-producing factors may prove synergistic and more physiologic. In fact, Moon and coworkers
18
have already reported that the combined transfer of Transforming growth factor Beta-1 (TGF-b1), Insulin like growth factor(IGF)-1 and BMP-2 revealed the aforementioned hypothetical synergistic effect of these factors on protein synthesis by demonstrating an amplification of protein synthesis.
An opposite approach that might be a potential alternative to the use of anabolic factors to induce disc cells into the production of matrix com­ponents is the application of anticatabolic factors. Inhibition of catabolic activity would ensue in maintaining or increasing the content of the respec­tive matrix component by slowing down its degradation without the need to force the disc cells to higher synthesis rates. Wallach and colleagues
19
recently published an in vitro study utilizing an adenoviral vector to introduce the gene encoding forTissue inhibitor of metalloproteinase (TIMP)-1 into disc cells isolated from degenerated human IVD. Gene delivery of TIMP-1 increased the proteoglycan content in the disc cell cultures, suggesting the anticatabolic approach to be a potentially promising strategy for gene therapy of degenerative disc disease.
The aforementioned index gene therapeutic approaches report results that sound promising. Despite the obvious potentials, the reality is that the application to human IVD will be challenged by the suboptimal and eventually toxic microenvironment inside the severely degenerated disc. It is questionable if the remaining compromised IVD cells in the degenerated disc will be able to produce reasonable amounts of gene-induced growth factors over extended periods of time. Furthermore, one can argue that it is unlikely that the existing starving cells are able to properly respond and produce an improved matrix even if the production of the respective gene product is achieved.
C H A P T E R   6 3     Intradiscal Biologics: A Potential Minimally Invasive Cure for Symptomatic Degenerative Disc Disease?
Autologous Implantation of Cultivated, Modified Cells
Degenerated discs could be treated by supplementation of the deserted matrix with in vitro cells that have been removed, cultivated, and modified. Autologous cells are optimal because their utilization makes the potential immunological complications a moot issue. Autologous cells compatible with disc tissue have to be harvested, expanded in vitro, and subsequently implanted into the symptomatic IVD. Once the cells have been removed and cultivated in vitro, this approach allows for indirect gene therapy via genetic modification of the withdrawn cells and/or tissue engineering via seeding of the cells in supporting biomaterials before implantation into the symptomatic degenerated IVD (see Figure 63-5). The combination of these techniques potentially improves efficiency by improving cell survival or enhancing the biosynthetic activity of the implanted cells. Genetic modi­fication of cultivated cells in vitro is technically very similar to the aforemen­tioned approaches previously discussed. The focus in this section turns to the cultivation of disc cells and creation of suitable implants.
For several reasons, it is extremely difficult to obtain suitable and suf­ficient numbers of target cells from intervertebral disc tissue. Removal of nucleus pulposus cells, the obvious target cells, would require opening of the annulus fibrosus to gain access. This would almost certainly cause damage to the annulus. In addition to the very restricted accessibility, the very low cell density in degenerated discs will further complicate the acquisition of ample usable cells for successful in vitro cultivation. Therefore only a limited number of scenarios are conceivable that would allow the withdrawal of suf­ficient cells from the disc to perform a disc cell–based approach without further damaging the already affected disc or accelerating its degeneration. Withdrawal of herniated disc material might be one scenario that would facilitate the removal of sufficient disc cells for in vitro cultivation. How­ever, the introduction of cells/implants after a surgical intervention on the same disc could be disputed since the outcome of microdiscectomy has been shown to be satisfactory for most patients with radiculopathy. That being said, the direct insult as well as the likely accelerated degenerative process of the surgical level post microdiscectomy predisposes that segment to dis­cogenic LBP postoperatively. This increased risk may justify cell implanta­tion to prevent postoperative acute and/or chronic discogenic LBP. Another potential clinical application would be the use of cell-based approaches to prevent the accelerated degeneration of discs adjacent to an interbody fusion level. Mechanical stress at segmental levels adjacent to fusion procedures is a known biomechanical issue. This is known to result in accelerated rates of degenerative disc disease and consequently discogenic low back pain at segments juxtapositioned to the fusion level (Figure 63-8). The disc mate­rial removed during the fusion procedure could be used as a source of cells to treat the adjacent disc. However, this would imply an intervention at an asymptomatic nondegenerated disc that only has the potential to degenerate in the future and is therefore rather questionable. Thus, at this time, autolo­gous disc cell transplantation is limited to a few clinical scenarios but has the potential to prove to be a powerful approach within these limitations.
The most direct approach to support a degenerated disc by autologous cells would be injecting a suspension of ex vivo proliferated disc cells. An approach to prepare autologous disc cells for subsequent transplanta­tion into the degenerated disc is the cultivation of the disc cells in three­dimensional cultivation systems. Initial experiments by Maldonado and associates constructs conserved the native phenotype, as demonstrated by the synthe­sis of matrix components similar to those observed in native discs. Since then, a wide variety of techniques have been recently applied to supply disc cells with the desired three-dimensional contructs. gies whose feasibility has been tested in vivo in animal experiments have also been undertaken. Gruber and colleagues, rat–based model, applied autologous disc cells, expanded in routine mono­layer cultures and seeded into a three-dimensional scaffold, to a hollowed cavity created in an intervertebral disc. After up to 33 weeks, no giant cell response was observed and the cells showed an appropriate morphology. In addition, no abnormality in the cell-surrounding matrix was observed, suggesting appropriate survival of the implanted cells during the analyzed time period. From their data the authors concluded that autologous disc cell implantation can be successful, although technically challenging. However, because of the immediate implantation after the seeding of the scaffold, the
21
demonstrated that cultivation of disc cells in three-dimensional
22
Experimental strate-
23
in a study utilizing a sand
F IG UR E 6 3 - 8  Lateral plain x-ray  demonstrating adjacent  level loss of 
disc height and degenerative changes at L4-L5 status post instrumented poste­rior lumbar interbody fusion at L5-S1.
disc cells do not have the time to synthesize appropriate amounts of matrix before encountering the adverse environment within the disc. Cultivation of the disc cells in a three-dimensional system before implantation might, therefore, improve the chances for survival in the hostile environment of the degenerated disc. Sato and coworkers
24
evaluated this approach by utiliz­ing a nonimmunogenic atelocollagen scaffold to seed and cultivate annulus fibrosus cells. This process demonstrated an increased ability to express type II collagen mRNA and deposited more type II collagen and proteoglycan compared to cells grown in monolayers. Atelocollagen scaffolds seeded with annulus fibrosus cells have been allografted into the lacunae of recipient rab­bits after laser diskectomy of the nucleus pulposus. Implantation resulted in a significant prevention of the narrowing of the intervertebral disc space up to 12 weeks postoperative compared to the nucleotomized control ani­mals. Histological analysis also showed that the allografted cells were via­ble, proliferated, and produced a hyaline-like matrix in the disc tissue of the recipients. Although the rabbit model does not appropriately simulate
20
the mechanical forces experienced by implanted disc cell in human discs, it is conceivable that cells surrounded by their own matrix might withstand mechanical forces with more success. That being said, another problem not addressed by the presented studies is the acute shortage of nutrients experi­enced by the cells after implantation into the degenerated disc. Considering that the nutrient supply is hardly sufficient for the original disc cells, it is questionable if the additional cells will survive for the prolonged time span likely required to provide a sustained structural improvement of the disc.
Implantation of Mesenchymal Stem Cells
Adult mesenchymal stem cells (MSCs) are uncommitted pluripotent stem cells that are found in several tissues, such as skeletal muscle, bone marrow, synovial membranes, and the dermis. plasticity and have a high capacity of multilineage differentiation. Members of the BMP family of growth factors have been used thus far to induce dif­ferentiation of mesenchymal stem cells into chondrocytes. BMPs are not exclusively inducing cartilage differentiation, its expression needs to be carefully timed and modulated to prevent the subsequent gen­eration of bone structures. To overcome this problem, signal transduction
25,26
Mesenchymal stem cells are of high
27
423
However, since
424
P A R T V I I     Surgical Treatment Modalities: Lumbar Spine
and transcription factors, such as members of the Sox family as well as the Brachyury factor, that exclusively induce cartilage differentiation have been tested and demonstrated with encouraging results. that cocultivation of MSCs with disc cells might be sufficient to induce a disc cell–like phenotype in MSCs.
30,31
Although these data originate from
28,29
It has been found
in vitro experiments, it might be conceivable that MSCs would also differen­tiate in vivo after injection into the disc. Besides the cocultivation with disc cells, cultivating of mesenchymal stem cells in three-dimensional cultivat­ing systems appears to be sufficient to induce a nucleus pulposus–like phe-
32
notype.
Implantation of collagen gel–embedded mesenchymal stem cells into artificially degenerated rabbit discs resulted in preserved nuclear and annular structures, prevention of proteoglycan decrease from the nucleus pulposus, and increased disc height.
33,34
The implanted cells were shown to survive and express genetic markers typical for nucleus pulposus cells. Similar results were also found after injection of a bone-derived MSC sus­pension into rabbit discs and injection of gel embedded MSCs into rat coc­cygeal discs.
35,36
The use of mesenchymal stem cells provides a new and exciting approach to biologically treat disc degeneration with encouraging results thus far. The comparably easy access to autologous mesenchymal stem cells allows overcoming one of the major culprits of conventional approaches. The high expandability of MSCs together with the relative ease of harvesting the cells makes this approach highly attractive. However, continued and extended studies are required to assess the structure of the newly synthesized matrix with regard to biomechanical properties and prove its value under the mechanical loads the functioning spine must bear.

CONCLUSIONS

The mechanics of human gait confer constant and multiplanar loads to
the IVD. The progressive structural alterations to the IVD that occur in continuum with spine segmental degeneration are not benign. The func­tions of the disc require a mechanically stable structure with a highly spe­cialized matrix to confer the needed flexibility and physical strength to the spine. The known avascularity of the adult IVD restricts nutrient supply to diffusion and therefore poses a major challenge for the prolonged mainte­nance of the diskal matrix by its cellular components. The aforementioned mechanical stressors combined with the known inadequate nutrition even­tually creates a toxic environment, resulting in progressive destruction of the matrix cellular structure and simultaneous extensive decay of the matrix. These properties and its alterations during degeneration define and limit the techniques applicable to biologically repair degenerated IVD and create patient cures. This might indicate that the clinical application of intradiscal biologics to regenerate the structurally compromised IVD is in the distant future. Recent studies looking at various biological approaches to maintain and improve the structurally compromised IVD provide real leads into the exciting potentials of these novel treatments. Further basic science and clinical experiments both in vivo and in vitro are required to bridge the gap between scientific potential and clinical realities.

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