Добавил:
Sekretar
kiopkiopkiop18@yandex.ru
t.me/Prokururor I Вовсе не секретарь, но почту проверяю
Опубликованный материал нарушает ваши авторские права? Сообщите нам.
Вуз:
Предмет:
Файл:Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_531_Библиотеки_им_академика_М_И_Перельмана
.pdf
ments and models. Eur J Cell Biol, 2006. 85(3-4): p. 165-73.
determine cell speed and direction. Mol Cell Biol, 2009. 29(6): p. 1506-14.
and gelsolin, in fetal skin explants. J Invest Dermatol, 2003. 120(6): p. 1118-29.
Nat Cell Biol, 2001. 3(5): p. E117-23.
glance. J Cell Sci, 2016. 129(6):1085-91
Life Sci, 2015. 72(16): p. 3051-67.
dynamics and traction forces in motile keratinocytes. FASEB J, 2016. 30(6):2298-310
dening form and driving function. Transl Res, 2011. 158(4): p. 181-96.
healing response of broblasts. J Cell Biol, 1993. 120(6): p. 1381-91.
wound healing reveals how cells mobilise back from the leading edge in a myosin-II-dependent
fashion. J Cell Sci, 2011. 124(Pt 7): p. 1017-21.
tiation in response to TGF-beta. Am J Physiol Lung Cell Mol Physiol, 2011. 301(5): p.
L656-66.
repair. Philos Trans R Soc Lond B Biol Sci, 2004. 359(1445): p. 777-84.
1(4): p. 1313-27.
and proliferation. J Pathol, 2007. 211(5): p. 572-81.
15(3): p. 133-39.
(Landmark Ed), 2011. 16: p. 1536-50.
The Role of Actin Remodelling Proteins in Wound Healing and Tissue Regeneration
http://dx.doi.org/10.5772/64673
153
and wound healing (review). Int J Mol Med, 2014. 34(1): p. 3-12.
ERM proteins, and signaling molecules during quiescence and wound healing. Histochem Cell
Biol, 2004. 121(5): p. 361-69.
the control of cell motility. EMBO J, 2001. 20(11): p. 2723-41.
stellate cell migration and decreased brosis. Biochim Biophys Acta, 2008. 1782(9): p. 542-48.
proteins bind advanced glycation end products, an interaction that may play a role in the
development of diabetic complications. J Biol Chem, 2003. 278(28): p. 25783-89.
required for kindlin-mediated alphaIIbbeta3 integrin coactivation. J Biol Chem, 2012.
287(10): p. 6979-90.
Dermatol, 2011. 131(1): p. 245-56.
14): p. 2353-56.
at sites of cell adhesion. Cell Res, 2012. 22(11): p. 1533-45.
force transmission at cell-matrix adhesions. J Cell Sci, 2015. 128(7): p. 1316-26.
1512-14.
FERM ground. Trends Biochem Sci, 2004. 29(8): p. 429-35.
cellular adhesion and migration during wound repair. J Invest Dermatol, 2009. 129(8): p.
2031-45.
function. Cell Adh Migr, 2011. 5(2): p. 160-69.
Wound Healing: New insights into Ancient Challenges154
tion. Neuron, 2006. 52(5): p. 789-801.
and vinculin-containing cell adhesions. FASEB J, 2014. 28(1): p. 453-63.
J Physiol Cell Physiol, 2010. 298(2): p. C221-36.
deformation and maintenance of tension in the collagen matrix. J Invest Dermatol, 2015.
135(11): p. 2569-71.
upon EGF stimulation. J Mol Recognit, 2013. 26(11): p. 532-41.
and maintenance of tension in the collagen matrix. J Invest Dermatol, 2015. 135(11): p.
2852-61.
cytoskeleton in time and space. Physiol Rev, 2008. 88(1): p. 1-35.
and cell migration. Cell Adh Migr, 2011. 5(2): p. 181-86.
tropomyosin isoforms. Mol Biol Cell, 2003. 14(3): p. 1002-16.
of the TrkA receptor: implications for diabetic nephropathy. Diabetes, 2012. 61(9): p. 2280-88.
Dermatol, 2013.
Cell Biol, 1999. 11(1): p. 103-8.
binding proteins as novel transcriptional coactivators. Bioessays, 2005. 27(4): p. 388-96.
leucine-rich repeats and gelsolin-like domains. Biochem Biophys Res Commun, 1999. 257(1):
p. 111-16.
bridging the leucine-rich repeat and the gelsolin superfamilies. J Biol Chem, 1998. 273(14): p.
7920-27.
274(47): p. 33179-82.
The Role of Actin Remodelling Proteins in Wound Healing and Tissue Regeneration
http://dx.doi.org/10.5772/64673
155
unique role for CapG in receptor-mediated ruing, phagocytosis, and vesicle rocketing. J Cell
Biol, 2001. 154(4): p. 775-84.
and leucine-rich-repeat family member: overlap with LLGL. Genomics, 1997. 42(1): p. 46-54.
Life Sci, 2004. 61(19-20): p. 2614-23.
mediated signalling. Biochem Soc Trans, 2004. 32(Pt 6): p. 940-42.
cell contraction. Exp Cell Res, 1999. 250(1): p. 155-67.
The Australian Journal of Wound Management, 2006. 14: p. 39-42.
broblasts that overexpress gelsolin. Science, 1991. 251(4998): p. 1233-36.
regulates cell migration. FASEB J, 2012. 26(8): p. 3260-72.
phosphorylation via a Rac1-dependent pathway. J Invest Dermatol, 2011. 131(7): p. 1450-59.
leucine-rich repeat Fli-I-interacting proteins. J Immunol, 2009. 182(6): p. 3450-60.
dependent transcription. Nucleic Acids Res, 2006. 34(18): p. 5052-59.
dampens cytokine secretion. J Cell Sci, 2012. 125(Pt 18): p. 4288-96.
inammation. Commun Integr Biol, 2012. 5(6): p. 546-49.
wounds and wound uid: neutralizing its activity in chronic but not acute wound uid improves
cellular proliferation. Eur J Dermatol, 2012. 22(6): p. 740-50.
modulating intracellular localization and caspase activity. J Cell Biol, 2008. 181(2): p. 321-33.
mammalian embryonic development. Mol Cell Biol, 2002. 22(10): p. 3518-26.
Wound Healing: New insights into Ancient Challenges156
gene. Genesis, 2011. 49(8): p. 681-88.
mental regulator, in early-gestation fetal wounds improves healing. Mamm Genome, 2011.
22(5-6): p. 341-52.
recovery following skin blistering. J Pathol, 2014. 232(5): p. 541-52.
Dermatol, 2015. 24(9): p. 692-97.
FLII in regulating Rac1-driven cell migration. Nat Commun, 2016. 7: p. 10664.
genesis in a murine model of type 1 diabetes. Diabetologia, 2014. 57(2): p. 402-12.
blistered skin in a murine model of epidermolysis Bullosa Acquisita. J Invest Dermatol, 2012.
133(4):1008-16
studies. Wound Repair Regen, 2012. 20(4): p. 523-36.
autoimmune ColVII mouse model of epidermolysis bullosa acquisita. J Pathol, 2011. 225(3): p.
401-13.
hypertrophic scarring and a target for a novel anti-scarring therapy. Br J Dermatol, 2015.
225(3):401-13
The Role of Actin Remodelling Proteins in Wound Healing and Tissue Regeneration
http://dx.doi.org/10.5772/64673
157
Chapter 7
Provisional chapter
Regenerative Approaches in Wound Healing: New
Alternatives for Older Tools
Michele R Colonna, Lupo Flavia, Delia Gabriele and
Stagno d’Alcontres Francesco
Additional information is available at the end of the chapter
Abstract
Critical wounds are well known to develop in elderly people and in other conditions
where inammation, vascular, and nervous disease lead to chronical ineciency in
running up healing processes. Recent researches have been focusing on microenviron‐
ment, and specic technologies have contributed to design and produce new materials
(the era of biomaterials and devices in wound healing).
At present, regenerative medicine and surgery have introduced a new approach, based
on cells’ transplantation producing specic cytokines and stimulating healing. This is
the role played by fat transplantation combining both stromal and vascular cellular
progenitor lines, monocyte products, platelet‐rich plasma, and glues.
New tools, such as templates and materials (comprising micro‐ and nanoparticles), as
well as technologies, such as cell seeding and gene therapy, revealed promising in this
direction. The authors report their experimental evidences and clinical experiences with
lipografting and peripheral blood mononuclear cells in critical wounds, focusing on
how to treat monocyte/macrophage cell depletion as well as insucient vascular supply.
Keywords: adipose‐derived stem cells, angiogenesis, dicult wounds, lipografting,
monocytes, vascular supply
1. Introduction
Wound healing is a highly structured physiological process involving cells and signal
molecules; it is known to run throughout inammation, cell proliferation, angiogenesis,
Regenerative Approaches in Wound Healing: New
Alternatives for Older Tools
Michele R Colonna, Lupo Flavia, Delia Gabriele and
Stagno d’Alcontres Francesco
Additional information is available at the end of the chapter
http://dx.doi.org/10.5772/64133
collagen deposition, and re‐epithelization [1]. In the very rst steps, natural immunity [2] plays
an important role through aggregation of inammasomes. The next step goes through
inammatory cell types, and messages from the rst to this step together with monocyte
responsivity are able to determine whether inammation will prolong to a sort of a steady,
chronically established state, which freezes the whole process transforming it into a chronic
inammation [2–6].
Monocyte/macrophage sequestration together with their lack of switch to type 2 [3, 4, 6]
impairs angiogenesis and cell activities resulting in a delayed re‐epithelialization, reduced call
for broblasts and diminishing collagen deposition, as well as a decreased cell proliferation.
The aim of this chapter is to report our experience on how to modify impaired wound healing,
starting from our experimental studies and concluding with our recent clinical experiences.
2. Pathophysiology
Following our studies [2] and those by Mirza et al. [4, 5], we started to focus on the role of
accumulation and insuciency of macrophages in wound healing. Macrophage dysfunction
has been shown to produce prolonged inammatory responses in critical wounds, in diabetes
through local secretion of proinammatory cytokines, such as IL‐1β, TNF‐α, MMP‐9, and IL‐6,
in particular, lack of switch to a second macrophage phenotype secreting proregenerative
factors, such as IL‐10, IGF‐1, and TGF‐β [6].
The result is an important interruption of healing with persistence of inammation, whether
hyperglycemia through cascade of ROS is its cause or not.
This produces critical wounds. Obviously, single biological steps can be analyzed in experi‐
mental studies, whereas in clinical practice this single factor must often be added to other
associated diseases, such as reduction of blood supply due to vascular obstruction. In the laer
case, translation from experimental to clinical work must also take into account adding
proangiogenetic factors even to provide a complete care.
Our clinical experience deals with persistence of inammation through both monocyte and
vascular insuciency.
2.1. Our experimental studies on wound healing
In this section, we report our experimental evidences leading to the results about the above‐
mentioned inammasome inhibition [2]. Plastic surgeons together with pharmacologists of
the University of Messina have been working on wound healing for more than 15 years
developing some models on rodents.
2.1.1. Animals
For these experiments adult male mice have been chosen, caged alone, and maintained under
a controlled environment (12‐hour light cycles day/night and 23°C room temperature, food,
Wound Healing: New insights into Ancient Challenges160
and water ad libitum) following Helsinki’s declaration and European and Italian laws and
Guidelines for Animal Laboratory Experiments.
Animals aging 8 weeks and weighing 20–25 g were used.
2.2. Model of incisional wound healing
The animals (mice) underwent general anesthesia with sodium thiopental (80 mg/kg intra‐
peritoneal injection) were scrubbed with iodine povidone on their back – their back skin shaved
and rinsed with physiological serum. Two longitudinal parallel 4 cm incisions were performed
and sutured with alternate stitches placed 1 cm from one another.
Sacrices were conducted on Days 7 and 14, respectively, and wounds were divided into three
segments of 80 mm large and 120 mm long. Caudal and cranial strips were used for molecular
studies, whereas the central one was used for histology and immunohistochemistry.
2.3. Model of dorsal skin ap
“Double H” aps were described following Quirinia et al.’s classication: this model was
modied and adopted [7, 8].
The animals (mice) underwent general anesthesia with sodium thiopental (80 mg/kg intra‐
peritoneal injection) were scrubbed with iodine povidone on their back – their back skin shaved
and rinsed with physiological serum. Double H ap consists of two opposite aps: the former
pedicled from the cranial side and the laer from the caudal one, both to be incised and elevated
on the dorsal skin. The central horizontal wound together with the two distal parts of the aps
acts as critical ischemia‐risk area. The aps were sutured back with separate stitches in Nylon
4/0. The animals were divided into three groups and sacrices were performed on Days 3, 6,
and 12, respectively. As for incisional wounds, three ap segments were taken and processed
the cranial and distal one for molecular essays, the central one for histology and immunocy‐
tochemistry
2.4. Model of burn injury
Under the same anesthesia and dorsum preparation described above, the dorsum of mice was
immersed in an 80°C bath for 10 seconds to produce a scald burn. Fluid resuscitation was
achieved through a subcutaneous 1 mL saline injection, treated with dierent agents (last used
EPO Z in comparison with EPO alpha), divided into three groups, and sacriced on Days 3,
6, and 12, respectively. Burned skin has been divided into two aps that have been, respectively,
used for molecular essays and histology.
Molecular essays are consisted in cytoplasmic protein dosage (Bio‐Rad Protein Assay (Bio‐Rad
Lab, Richmond, CA, USA), spectrophotometry, using albumin as a standard); Western blot for
GFs and cell cycle molecules; histology measured the presence of inammatory inltrates,
necrosis, and repair in standard hematoxylin (eosin, trichromic, and immunohistochemistry
were used to visualize and quantify alpha‐smooth positive cells such as a response to VEGF
in both neoangiogenesis and neovasculogenesis).
Regenerative Approaches in Wound Healing: New Alternatives for Older Tools
http://dx.doi.org/10.5772/64133
161
Statistical analysis was conducted with parametric essays for repeated measures (ANOVA)
and bonferroni test was used to evaluate intergroup positivity, with a p ¼ 0.05 considered as
statistically signicant.
Our experimental studies have pointed out some important features of antioxidant molecules
in impaired wound healing (diabetic mice), as well as the role of some cytokine‐related
molecules and endogenous products belonging to natural immunity cascade [2] in normal and
impaired wound healing [9–13].
2.5. Another important contribution to the study of neoangiogenesis and biomaterials
prefabrication
Our group also developed a collaboration with the group from Padriciano, International
Center for Genetic Engineering and Biology, United Nations, to study a model of prefabricated
ap in the groin of adult rats creating an artero‐venus loop that was included into a dermal
regenerative template; this new regeneration chamber was then injected with dierent viral
vectors (AAV 156) encoding for the production of VEGF. Results were remarkable, demon‐
strating enhancement of neoangiogenesis and neovasculogenesis and the utility of this novel
model of regeneration chamber that could act as a bioreactor and stimulate healing and even
repair poor vascularized tissues [14, 15].
3. Clinical experiences
Chronic wounds are produced either by an interruption in healing processes, as an eect of
lack of positive (vascular supply and neurotrophism) or of an excess of inhibitory factors
(metallo proteinases in ECM, some cytokines), or by a lack of switch in inammatory cell
phenotypes, such as in diabetes.
As a nal eect, wound bed does not progress beyond detersion, typically presenting itself as
necrosis or debris. They are especially present in lower limbs, often as a result of complex mix
of the above‐mentioned factors.
Vascular and diabetic ulcers are the most common chronic wounds aecting nearly 2–5% of
the general population and have received an important impact in terms of morbidity, absence
from work, and social costs.
Traditional wound dressings do not restore vascular supply, which is a sine qua non for
restarting healing.
An important role is played by vascular surgery and endovascular techniques, which act
restoring the lost vascular supply or producing bypasses to revascularize the area.
At present, some novel suggestions come from regenerative surgery.
Wound Healing: New insights into Ancient Challenges162
Соседние файлы в папке Библиотека им академика М.И. Перельмана