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Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_5667_Библиотеки_им_академика_М_И_Перельмана

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 
n
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226
Figure8.2 Hyaluronic acid benzyl ester.
degrees of esterification, typically performed with benzyl alcohol, have been experi-
®
mented with. HYAFF
is a registered HA benzyl ester (see Figure8.2), employed in
particular for wound healing[29] and cartilage tissue repair[30].
Cutaneous lesions generated by burns, ulcers, or trauma are usually cured with skin grafts; however, if the covering process cannot be immediately performed, biomaterialbased patches are employed to induce the correct recovery and regen­eration of the damaged area[31]. HYAFFbased medical devices have been success­fully employed for the regeneration of cutaneous lesions, and the different esterification degrees and formulation methodologies allow for modulation of the final shape/geometries of the device[32]. HYAFF has also been employed in chon­drogenic tissue engineering applications for its ability to induce cartilage forma­tion[33]. In vitro studies employing 3D HYAFFbased matrices showed the ability of autologous chondrocytes to restore the physiological signatures of hyaline carti­lage[34], whereas invivo studies demonstrated that seeding with autologous chon­drocytes induces native cartilage tissue formation with optimal integration in the surrounding tissue[33]. Hyalograft C, a medical device based on cartilage autograft obtained from patient chondrocytes cultured on HYAFF, was introduced for surgi­cal arthroscopic techniques in 1999 [35, 36]. In 2013, following an European Medicines Agency (EMA) report on manufacturing practices, Hyalograft C was withdrawn from the European market[37]. Today, methodologies for treatment of cartilage include new approaches in which autologous chondrocytes are delivered into damaged tissues with HA or collagenbased injectable biomaterials[38]. HA is also employed, in different formulations, in ophthalmic viscoelastic devices (OVDs) or dermatological therapies[39]. The first use of untreated HA in ophthalmic sur­gery was reported in cornea transplantation and, today, it is currently employed in ocular surgeries for corneal protection and vitreous replacement [40]. Food and Drug Administration (FDA) approved the first viscosurgical device based on HA in
®
1983, under the trademark Healon
 [41, 42]. Healaflow®, an FDAapproved HA crosslinked with 1,4butanediol diglycidyl ether (BDDE), is employed in vitreoreti­nal surgery[43]. Other ophthalmic applications include the modification of contact lenses with HA solutions. Recently, HA has been exploited to improve contact lens performance or obtain a controlled release of HA in the eye[44, 45]. Other FDA approved products based on crosslinked HA include hydrogels in which the crosslinking is performed with divinyl sulphone (DVS), such as commercial dermal
8.2 Biomaterials and Medical Devices: Natural and Synthetic Strategies
defect fillers (Hylaform®, CaptiqueTM, Prevelle®, Lift®, and Varioderm®). In the cosmetic field, the most employed HAcrosslinked hydrogels are based on the men-
®
tioned HADVS and HABDDE (Restylane
, Juvéderm®, Teosyal®, and Hyabell®) as
injectable materials[39, 46–48].
Sulfated polysaccharides, normally linked to proteins as PGs, have impressive bio­chemical and structural roles in the regulation of tissue morphology and cell devel­opment and have consequently been extensively employed in medical devices and studied for clinical translation. Among them, CS, a sulfated linear polysaccharide based on repeating units of glucuronic acid (GlcA) and Nacetylgalactosamine (GalNAc) (Figure 8.1), has been largely employed in the biomaterials field [2, 49–51]. The sulfation pattern of CS, usually at the positions4 and 6, affects the physiological role. CS is found in many tissues and organs of the human body, from central nervous system (CNS) to cartilage, skin, bones, and blood vessels. In particu­lar, the major efforts in the development of CSbased medical devices are focused on the replacement or treatment of cartilage defects and the regeneration of CNS func­tions. CS is often employed in combination with other natural polymers to generate
®
hybrid medical devices. Esoxx
is a hybrid medical device employed as mucosal bar-
rier for the treatment of gastroesophageal reflux symptoms based on HA and CS
®
embedded in Lutrol
F 127, a poloxamer with bioadhesive properties [52]. HA/
CSbased medical devices are also employed as injectable hydrogel to treat osteoar-
®
thritic (OA) knee, as reported for Structovial CS
[53]. In this formulation, CS has a double role: optimizing the HA rheological properties and regulating the cartilage metabolism and remodeling[53]. CS is also extensively employed in formulations with glucosamine for oral administration as Symptomatic Slow Acting Drugs for Osteoarthritis (SYSADOA), even if this use is still debated and several clinical stud­ies are ongoing[54]. Other sulfated polysaccharides exploited in clinical translation are HS and heparin (Figure8.1). HS is heavily expressed in the ECM and on the cell surface as HSPGs . The structure and length of the polysaccharide chains are highly variable due to the different sulfation degrees, the different combinations of disac­charide components, and, like for other ECM polysaccharides, the variable molecu­lar weight [55]. The typical disaccharide components consist of 40–60% of βglucuronic acid (GlcA) or αiduronic acid (IdoA) linked by (1 → 4) glycosidic bonds to glucosamine, which can be Nacetylated or Nsulfated[56, 57]. Mimetics of HS have been developed by derivatizing dextran with carboxymethyl, carboxym­ethyl benzylamide, and carboxymethyl benzylamide sulfonate groups[58, 59]. An
®
example is a commercially available product named RGTA the treatment of chronic skin damages (CACIPLIQ20
®
(Cacicol20
). Other applications of RGTA are under study and include osteochon-
, currently employed for
®
) and for corneal regeneration
dral, gastroenterological, cardiac, and nervous system applications[60]. Heparin contains the same monosaccharides of HS, which can also be, in this case, sulfated as Ido(2S) and GlcA(2S), and the GlcN unit can be Nsulfated (GlcNS) or N acetylated (GlcNAc) with differential patterns and degrees[61]. The substitution patterns with Osulfate and Nacetyl groups result in a wide range of complex structures. Also, heparin has been largely investigated and employed in the pharmaceutical and
227
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228
material device fields for different applications, even if, considering its main role as an anticoagulant, the use of heparinbased medical devices is mainly focused on bloodcontacting materials[61, 62]. Temporary or permanent devices like hemodi­alysis catheters, coronary or vascular stents and grafts, bypass devices, and extracor­poreal circulation devices exploit heparincoated materials[57, 62–64]. The two general methods employed for the development of heparincoated devices are based on eluting technologies and noneluting technologies. Eluting technologies include all the approaches in which heparin is released in a controlled way, taking advan­tage of noncovalent coatings based on physical or ionic interactions [62, 65, 66]. These methods are employed when heparin delivery is needed to prevent acute and local devicerelated blood clots. The other approach is based on the covalent func­tionalization of the medical device surface with heparin[67, 68]. This approach is desirable when longterm thromboresistance is needed at the material surface. Different conjugation methodologies exploit the direct coupling of complementary functional groups already present in heparin and the material surface counterpart, or alternatively introduce new chemoselective orthogonal functional groups and properly tailored linkers to better control heparin exposition and consequently the antithrombotic activity. The chemical nature of the medical device will influence the choice in light of the functional groups eventually present at the surface or, alternatively, introducible upon proper treatment. The most employed conjugation methodologies are based on carbodiimide chemistry, exploiting the carboxyl group of heparin, or oxidation mediated by periodate, for controlled oxidative degradation to generate some aldehydes[69]. A coating material based on heparin, polyethylene oxide, and sulphonate groups covalently conjugated to hydrophilic modified poly-
TM
ethyleneimine (ASTUTE and licensed to Medtronic as Trillium dialysis catheters[62, 64]. Another example is Bioline
technology) has been developed by BioInteractions Ltd
®
to cover cardiopulmonary bypass and hemo-
®
, developed by Jostra AG and now a property of Maquet Medical, a material obtained with heparin linked to human albumin employed for coating vascular grafts and cardiopulmonary bypass[63, 70]. The conjugation of heparin to polyamine carriers, performed by introducing etherofunctional crosslinkers, is currently employed as Corline Heparine Surface Technology, developed by Corline System AB. Many other coating methodologies employed for the modification of medical device surfaces to control the interface between blood components and material surface, and to minimize blood clot formation and thrombolytic events are reported with other commercial examples in Table8.1.
®
8.2.1.2 Polysaccharides fromPlants, Algae, Animal, and
Microbial Fermentation
The large availability of polysaccharides derived from plants, algae, and animals and the possibility to produce them by microbial fermentation make them very attractive and cheaper starting materials[13]. Several studies are focused on the biocompatibility of such polysaccharides, and many of them are already in clinical applications. These polysaccharides present an interesting variety of biochemical and physical properties that can eventually be advantageously exploited in
8.2 Biomaterials and Medical Devices: Natural and Synthetic Strategies
Table8.1 Examples of commercial heparin-coating technologies.
Polymer Commercial names Application References
229
HA benzyl ester
Sodium hyaluronate
HA–BDDE
HA–DVS and HA–BDDE
HA–CS embedded in
®
Lutrol
HA–CS
Mimetics of HS–derivatized dextran with sulfonate groups
Heparin, polyethylene oxide, and sulfonate groups covalently conjugated to polyethyleneimine
Heparin linked to human albumin
Heparin linked by reductive amination to matrices bearing amino groups
Heparin UV photocrosslinked
Layer by layer coatings. The last layer (4) is coated with polyethyleneimine linked to Heparin by reductive amination
Heparin and benzalkonium chloride blended
HYAFF
®
Osteochondral regeneration, wound
[29, 30, 32–34]
healing
®
Healon
Healaflow
®
HADVS (Hylaform
TM
Captique Varioderm (Restylane Teosyal
Esoxx
, Prevelle®, Lift®,
®
) HABDDE
®
, Juvéderm®,
®
, Hyabell®)
®
®
,
Viscosurgical devices [41, 42]
Vitreoretinal
[43]
surgery[43] Dermal defect fillers [39, 46–48]
Treatment of gastro
[52] esophageal reflux symptoms
Structovial CS
®
Treatment of osteoarthritic (OA)
[53]
knee
®
RGTA
(CACIPLIQ20® and
®
Cacicol20
)
ASTUTE™/TRILLIUM coating
BIOLINE
®
Coating
Chronic skin damages and corneal regeneration
Hemodialysis
®
catheters, bypass devices
Vascular grafts, extracorporeal
[58–60]
[62, 64]
[63, 70]
circulation devices
CARMEDA
®
Surface (CBAS Surface)
BioActive
®
Heparin
Extracorporeal circulation devices, stentgrafts, vascular grafts, ventricular
[62, 71]
assist devices
PHOTOLINK
®
Various medical devices
[62]
Hepamed™ Coronary stents [62, 72]
DURAFLO II
®
Extracorporeal circulation devices
[62]
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230
biomedical applications. Furthermore, they can be properly modified to control the properties and final morphologies of a large plethora of formulations. The main nonhuman polysaccharides exploited for biomedical devices are alginates, carra­geenan, and fucoidan (derived from algae), dextran, pullulan, and gellan (derived from microbes), and chitosan (derived from animals) (see Figure8.3).
–
+
COO
O
HO
HO
G
Na+ –OCC
O
HO
R
2
O
OH
O
OH
HO
OH
1
O
HO
O
OH
Alginate
OH
HO
O
O
NH
2
Chitosan
OH
O
O
R
1
Carrageenan
R= –OH, OSO
O
O
OH
OH
O
HO
Pullulan
OH
O
HO
R
OH
M
O
OH
5
R
O
–
3
OH
Na
NH
2
O
O
n
OH
3
O
O
R
4
n
O
OH
O
O HO
HO
O
HO
OH
O
O
HO
n
HOOC
O
OH
O
HO
O
HO
Gellan gum
HO
HO
OH
HO
HO
OH
O
HO
OH
O
O
OH
O
O
O
OH
HO
Dextran
OH
Fucoidan
O
OH
OH
O
OH
O
O
HO
OH
O
HO
OSO
OH
OH
O
OH
O
HO
O
O
OSO
O
O
–
OSO
3
– 3
HO
O
O
O
O
O
n
n
O
O
–
OSO
3
–
3
Figure8.3 Examples of nonhuman polysaccharides employed for biomedical device formulations.
8.2 Biomaterials and Medical Devices: Natural and Synthetic Strategies
Alginates are linear polysaccharides obtained from seaweed composed of β mannuronate (M) and αguluronate (G) residues linked by (1,4) glycosidic bond[73]. The polysaccharide structures are organized in blocks with consecutive M (MMMMMM) or G (GGGGGG) residues or alternating M and G residues (GMGMGM), depending on the algae from which they are extracted. The hydrogel properties of alginates are related to the M/G ratio and sequence, the Gblock length, and finally the molecular weight[74]. Alginates are widely investigated for biomedi­cal applications thanks to their biocompatibility and the ability to form hydrogels by
2+
simple addition of divalent cations like Ca
, and several of them have found appli-
cations even in clinics. Alginate dressings have been experimented in wound heal-
TM
ing, and are commercially available as Algicell
, AlgiSite M™, Comfeel Plus™, Kaltostat™, Sorbsan™, and Tegagen™; they show an improved ability to support rapid healing through epithelialization[75]. Alginatebased biomaterials are also employed for dental impression applications[76] and for the treatment of gastro esophageal reflux [77, 78]. Other interesting biopolymers derived from algae are carrageenan and fucoidan. Both are sulfated polysaccharides, derived, respectively, from red and brown algae[79]. Carrageenans are sulfate galactans composed of  galactose (Gal) and 3,6anhydrogalactose with alternating α(1→3) and β(1→4) glycosidic bonds; depending on the structure and sulfation patterns, they are classi­fied as γ, β, δ, α, μ, κ, λ, ν, ι and θcarrageenans. Carrageenans are currently employed as food additives and nutraceuticals; however, several studies highlight their potential use in drugdelivery systems, tissue engineering, and wound heal­ing[80]. Fucoidans are highly sulfated polysaccharides based on repeating units of α(1→3)fucopyranose, often alternated with α(1→4)fucopyranose units. They can also contain acetate groups and various glycosyl side branches (i.e. glucuronic acid, mannose, glucose, galactose, or xylose), depending on the type of brown algae. Like carrageenan, fucoidan is currently employed in the nutraceutical and food industry; however, several biomedical applications in the medical devices, drug delivery, and tissue engineering fields are under investigation.[81].
Polysaccharides derived from microbial sources are extensively studied for cos­metic, pharmaceutical, nutraceutical, and biomedical applications. Dextran is one of the most employed; it is α1,6polyαglucoside with α1,3 branches produced by microbial fermentation employing nonpathogenic Leuconostoc mesenteroides, Saccharomyces cerevisiae, Lactobacillus plantarum, and Lactobacillus sanfrancisco. Dextran is used in vascular surgery and antiplatelet applications thanks to its antithrombotic properties; furthermore, it finds application in ophthalmic and diag-
®
nostic applications. Dextran 70
(MMW 70.000 kDa) and Dextran 40® (LMW
40.000 kDa) are employed, respectively, as plasma substitutes and to increase blood flow in ischemic limbs[82, 83]. Lowmolecularweight dextran sulfate (LMWDS) is also under study (Phase II clinical trial) to reduce graft rejection for pancreatic islet transplantation[84]. Dextran is also extensively investigated as coating for diagnos­tic and therapeutic nanoparticles; in particular, dextrancoated magnetic nanoparti­cles are employed as diagnostic agents for MRI and as therapeutic nanoparticles for iron deficiency [85]. Other interesting examples of microbial polysaccharides include pullulan, employed for oral care and cosmetic applications[86], bacterial
231
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232
cellulose, employed for topical woundhealing applications[87], and gellan gum, employed in the development of oral, ophthalmic, and spray nasal formulations for biomedical, cosmetic, and nutraceutical applications[88, 89]. All the cited bacterial polysaccharides are also under investigation for other biomedical applications, including wound healing and regenerative medicine[88].
Chitosan is the most commonly used polysaccharide from animal sources; it is obtained from crustacean shell waste. Many chitosanbased biomaterials are com­mercially available, and some are in clinical trials for wound healing, pharmaceuti­cal, diagnostic, and cosmetic applications. Formulations and medical devices based on chitosan are usually obtained by ionic assembling with anionic polymers or by covalent linkages performed on the amino groups by carbodiimide chemistry. Another approach is based on the partial oxidation of chitosan with periodate generating aldehyde groups for subsequent chemoselective reactions (Table8.2).
Table8.2 Examples ofcommercial medical devices and formulations produced fromvegetal, microbial, or animal-derived polysaccharides.
Polysaccharide Commercial names Origin Application References
Alginate Algicell™, AlgiSite
Sodium alginate Neocolloi
Potassium alginate Blueprint Cremix®,
Sodium alginate Liquid Gaviscon
Carrageenan Carragelose
Dextran MMW Dextran 70 Dextran LMW Dextran 40 Dextran sulfate LMW Ibsolvmir Dextrancoated iron
nanoparticles
Dextrancoated iron nanoparticles
M™, Comfeel Plus™, Kaltostat, Sorbsan™, Tegagen™
®
, Zhermack Plus
Dentsply DeTrey
Endorem Combidex
CosmoFer, INFeD, Ferrisat, DexFerrum, and DexIron
®
; Palgat
®
, 3M ESPE
®
®
®
®
,
®
®
®
®
®
Algae Wound dressing [75]
Algae Dental impression [76]
Dental impression [76]
Algae Gastroesophageal
reflux
Algae Antiviral eye
drops, nasal spray Microbial Plasma substitute [82, 83] Microbial Plasma substitute [82, 83] Microbial Islet transplant [84] Microbial Imaging liver
cancer
(endoderm) and
lymph node
metastases
(Combidex) Microbial Iron deficiency
therapy
[77, 78]
[85]
[85]
8.2 Biomaterials and Medical Devices: Natural and Synthetic Strategies
Table8.2 (Continued)
Polysaccharide Commercial names Origin Application References
233
Pullulan Listerin Bacterial cellulose Bioprocess
Gellan Gum Grindsted Gellan Gum Gelrite
Chitosan HemCon
Chitosan and cellulose
Chitosan– glycerophosphate
Chitosandextran CD gel Animal
®
and Biofill
®
and Kelcogel others
Chitoflex
Chitoseal
BSTCarGel
©
® ®
®
Microbial Oral care [88]
®
, XCell®,
®
®
Gellan Microbial Nutraceutical [91]
many
,
, Celox
®
Microbial Wound healing [87, 90]
Microbial Drug release,
Animal Hemostatic
®
Animal and microbial
Animal Cartilage repair [93]
and microbial
ophthalmic, nasal formulations, cosmetic formulations
bleeding treatment and bandages
Bleeding wounds [93]
Surgical applications, wound healing
8.2.2 Carbohydrates asSignaling Molecules: Opportunities inTissue Engineering and Regenerative Medicine
[89, 92]
[93]
[94, 95]
Tissue engineering and regenerative medicine strategies are based on two main pil­lars: (i) biomaterial scaffolds mimicking the ECM and (ii) stem cells able to regener­ate tissues damaged from trauma or pathologies. The biomaterial scaffold can be implanted or just used to culture the human cells that will be implanted/adminis­tered subsequently. The role of the biomaterial scaffold in this case consists in hom­ing the cells and driving their fate to the required regeneration. In this context, glycans are deeply investigated not only for their structural role to generate cell hospitable hydrogels but also for their role as signaling molecules capable of modu­lating the cell’s fate. Recent studies have demonstrated that differential cell fate modulation is strictly dependent on the glycan epitopes present in the ECM[96, 97]. Considering that both stem cells and ECM are heavily glycosylated, it is easily understandable that a “glycocode” plays a key role in cellsignaling induction and transduction[4, 98]. Morphological and functional tissue morphogenesis, related to healthy and pathological states and even aging, is related to N and Oglycosylation of ECM proteins and cell–ECM communications[99]. Alteration in glycosylation of ECM and extracellular components has been characterized in pathological condi­tions like cancer[100–103], fibrosis[104, 105], and inflammatory diseases[106–108].
 
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234
Collagen, laminin, and fibronectin are the most known examples of ECM proteins in which glycosylation varies between physiological and pathological conditions and in tissue morphogenesis[23, 109, 110]. ECM–cell crosstalk events are mediated by carbohydrate–carbohydrate and, more significantly, by carbohydrate–protein (lectins) interactions[3, 111–113]. The crosstalk will result in different structural organizations of the ECM responsible for morphogenesis. Therefore, once the cor­relation between cell fate and ECM morphological and biomolecular properties (including glycosylation) is clarified, it will be possible to design artificial ECM able to regenerate the required tissue. Even if glycoproteomic and lectin studies are in rapid development, we still need a clear and reliable picture of the main glycosigna­ture changes related to morphogenesis and the occurrence of pathologies.
8.3 Carbohydrates inAnimal-Derived Medical
Devices: Friends or Foes?
Among implantable medical devices, animalderived organs or tissues are frequently the best choice for tissue substitution or surgical applications[114]. They not only provide the nanometrically correct architecture for the required functionality but also possess the biochemical composition ideal for cell adhesion and consequent tis­sue regeneration. As a matter of fact, even in synthetic medical devices, animal derived polymers are often employed as coatings to optimize the interactions between tissue and material surfaces or to control the medical device performances[115–117]. Bovine pericardium or bovine/porcine patches are employed for the reinforcement of shapes and lines in soft tissue surgery applications. Collagen and gelatinderived meshes and coatings, as well as bovine/porcinebased devices, are employed in wound dressing and vascular surgery applications. Animalderived dural grafts [118, 119] and nerve conducts[120, 121] are also employed in neurosurgery.
Animalderived materials, however, suffer from a big inconvenience: the presence of xenoantigens like αGal, Neu5Gc (N-Glycolylneuraminic acid), and Forssman antigen (GalNAcα1, 3GalNAcβ1, 3Galα1, 4Galβ1, and 4GlcCer) [122, 123] (see Figure8.4) that elicit an immune response in humans.
Strategies like genomic editing and chemical immunosuppression are under investigation to limit xenoantibody production and avoid undesirable early graft rejection[122, 124, 125]. In December 2020, transgenic pigs with depletion of α  Gal–named GalSafe pigs–were approved by FDA for human food consumption and possible biomedical uses [126]. Decellularized animal tissues were also developed with the aim to limit xenoantigens from cellular components; how­ever, the immunological response of these devices is often still in place, probably due to the glycosylation of ECM components[125]. The most efficient solution is probably a coating able to mask the xenoantigens responsible for immunore­sponse. Glutaraldehyde fixation [127, 128] is an option. Treatment of animal derived biological tissues with a low concentration of glutaraldehyde enhances structural performance, maintains sterility, and reduces antigenicity. Although
    
OH
HO
Forssman antigen
r
235
COOH
O
O
OH
HO
O
OH
O Ce
O
OH
HO
OH
OH
OH
α-Gal
NHAc
OH
HO
O
HO
O
OH
H N
HO
O
Neu5Gc
O
OH
OH
O
NHAc
OH
OH
O
O
OH
O
HO
O
OH
O
OH
Figure8.4 Xenoantigens responsible for immunoresponse in humans.
the advantages of this kind of coating are wellestablished, the calcification phe­nomena induced at the surfaces of the devices are still a remarkable limita­tion[129]. To reduce the risk of calcification, antimineralization treatments have been developed, such as the use of alphaamino oleic acid (AOA) employed in
®
Mosaic
bioprosthesis (Medtronic), which also improves durability and hemody­namic properties. The AOA reacts with the free aldehyde groups of glutaralde­hyde by Schiff base covalent linkages, avoiding calcium nucleation and precipitation[130]. Glutaraldehyde fixation is able to mask around 50% of αGal antigens[131], but at the same time, it increases oxidative and enzymatic degra­dation of the tissue. Recently, a new treatment available for glutaraldehydecoated
®
tissues has been patented (FACTA
), able to totally mask αGal epitopes in animalderived medical devices, reducing oxidative degradation and thrombotic risk [132].
8.4 Glycoengineering Application toRegenerative Medicine
Metabolic glycoengineering (MGE) approaches were developed in the late 1980s and are revealed today as valuable tools to study the expression of selected glycans in pathological phenomena and to modulate glycosignature expression [133–135].