PRODUCTION OF ACELLULAR DERMAL MATRIX OF PIG᾽S SKIN: MORPHOLOGICAL ANALYSIS
DOI:
https://doi.org/10.11603/1811-2471.2020.v.i4.11761Keywords:
pig᾽s skin, derma, acellular dermal matrix, morphologyAbstract
SUMMARY. The main propose on the treatment of skin defects of various etiologies is the use of decelularized (acellular) pig skin.
The aim – to create the method of production an acellular dermal matrix of the dermis of pig᾽s skin while preserving the native structure.
Material and Methods. The process of creating an acellular dermal matrix was a combination of physical and chemical effects on the dermis. Derma (1.0–1.3 mm thick) was taking from the back and partly from the lateral surfaces of the body of the 1 year – old pigs. Previously, a layer of skin 0.3–0.4 mm thick was removed from the relevant areas with the help of a dermatome. Achieving the maximum acellularization of the dermal matrix was carried out step by step and included 4 stages: 1 – the process of freezing-thawing; 2 – dehydration with glycerin; 3 – osmotic stress; 4 – washing with detergent.
Results and Discussion. Histological analysis of the dermis of pig᾽s skin after freeze-thaw cycles revealed that the collagen skeleton of the dermal matrix, obtained as a result of the first stage of decelularization, has normal structural organization.
The second stage of decelularization revealed a decrease in the number of fibroblasts. The connective tissue elements, which are represented by multidirectional bundles of collagen fibers, retained their structural organization.
The third stage of decelularization revealed the lysis of fibroblastic cells, which as the basophilic stained elements were localized in small quantities in the dermis.
Washing of lyophilized skin with nonionic detergent sodium dodecylsulfate revealed the complete absence of fibroblastic cells, epidermocytes in the hair follicles, endothelial cells in the wall of blood vessels, which indicates the effectiveness of this reagent in removing residual products.
Conclusions. The proposed protocol for deceleralization of the dermis of pig᾽s skin is effective in removing nuclear and cellular structures from the dermis. Specific protocols can be modified by increasing the temperature difference or changing the number of freeze-thaw cycles.
References
Debeer, S., Le Luduec, J.B., Kaiserlian, D., Laurent, P., Nicolas, J.F., Dubois, B., & Kanitakis, J. (2013). Comparative histology and immunohistochemistry of porcine versus human skin. Eur. J. Dermatol., 23 (4), 456-466.
Chua, A.W., Khoo, Y.C., Tan, B.K., Tan, K.C., Foo, C.L., & Chong, S.J. (2016). Skin tissue engineering advances in severe burns: review and therapeutic applications. Burns Trauma, 4, 3. DOI: 10.1186/s41038-016-0027-y.
Tracy, L.E., Minasian, R.A., & Caterson, E.J. (2016). Extracellular matrix and dermal fibroblast function in the healing wound. Adv. Wound Care (New Rochelle), 5, 119-136.
Chaudhari, A.A., Vig, K., Baganizi, D.R., Sahu, R., Dixit, S., Dennis, V., …, & Pillai, S.R. (2016). Future prospects for scaffolding methods and biomaterials in skin tissue engineering: a review. Int. J. Mol. Sci., 17 (12), 1974.
Sheikholeslam, M., Wright, M.E.E., Jeschke, M.G., Amini-Nik, S. (2018). Biomaterials for skin substitutes. Adv. Healthc. Mater., 7 (5), 1700897.
Cui, H., Chai, Y., & Yu, Y. (2019). Progress in developing decellularized bioscaffolds for enhancing skin construction. J. Biomed. Mater. Res. A, 107 (8), 1849-1859.
Hrebikova, H., Diaz, D., & Mokry, J. (2015). Chemical decellularization: a promising approach for preparation of extracellular matrix. Biomed. Pap. Med. Fac. Univ. Palacky Olomouc Czech. Repub., 159 (1), 12-17.
Crapo, P.M., Gilbert, T.W., & Badylak, S.F. (2011). An overview of tissue and whole organ decellularization processes. Biomaterials, 32 (12), 3233-3243.
Riau, A.K., Beuerman, R.W., Lim, L.S., & Mehta, J.S. (2010). Preservation, sterilization and de-epithelialization of human amniotic membrane for use in ocular surface reconstruction. Biomaterials, 31 (2), 216-225.
Olczyk, P., Mencner, L., & Komosinska-Vassev, K. (2014). The role of the extracellular matrix components in cutaneous wound healing. BioMed. Res. Int., 2014, 747584.
Volk, S.W., Iqbal, S.A., & Bayat, A. (2013). Interactions of the extracellular matrix and progenitor cells in cutaneous wound healing. Adv. Would Care, 2 (6), 261-272.
Xu, C.C., Chan, R.W., & Tirunagari, N.A. (2007). Biodegradable, acellular xenogeneic scaffold for regeneration of the vocal fold lamina propria. Tissue Eng.,13 (3), 551-566.
Cox, B., & Emili, A. (2006). Tissue subcellular fractionation and protein extraction for use in mass-spectrometry-based proteomics. Nat. Protoc., 1 (4), 1872-1878.
Xing, Q., Yates, K., Tahtinen, M., Shearier, E., Qian, Z., & Zhao, F. (2015). Decellularization of fibroblast cell sheets for natural extracellular matrix scaffold preparation. Tissue Engineering-Part C Methods., 21 (1), 77-87. DOI: 10.1089/ten.tec.2013.0666.
Prasertsung, I., Kanokpanont, S., Bunaprasert, T., Thanakit, V., & Damrongsakkul, S. (2008). Development of acellular dermis from porcine skin using periodic pressurized technique. J. Biomed. Mater. Res. B. Appl. Biomater., 85 (1), 210-219.
Levy, R.J., Vyavahare, N., Ogle, M., Ashworth, P., Bianco, R., & Schoen, F.J. (2003). Inhibition of cusp and aortic wall calcification in ethanol- and aluminum-treated bioprosthetic heart valves in sheep: background, mechanisms, and synergism. J. Heart Valve Dis., 12 (2), 209-216.
Gorschewsky, O., Klakow, A., Riechert, K., Pitzl, M., & Becker, R. (2005). Clinical comparison of the Tutoplast allograft and autologous patellar tendon (bone-patellar tendon-bone) for the reconstruction of the anterior cruciate ligament: 2- and 6-year results. Am. J. Sports Med., 33 (8), 1202-1209.
Gorschewsky, O., Puetz, A., Riechert, K., Klakow, A., & Becker, R. (2005). Quantitative analysis of biochemical characteristics of bone-patellar tendon-bone allografts. Biomed. Mater. Eng., 15 (6), 403-411.
Nakayama, K.H., Batchelder, C.A., Lee, C.I., & Tarantal, A.F. (2010). Decellularized rhesus monkey kidney as a three-dimensional scaffold for renal tissue engineering. Tissue Eng. Part A., 16 (7), 2207-2216.
Yang, B., Zhang, Y., Zhou, L., Sun, Z., Zheng, J., Chen, Y., & Dai, Y. (2010). Development of a porcine bladder acellular matrix with well-preserved extracellular bioactive factors for tissue engineering. Tissue Eng. Part. C Methods., 16 (5), 1201-1211.
Du, L., Wu, X., Pang, K., & Yang, Y. (2010). Histological evaluation and biomechanical characterisation of an acellular porcine cornea scaffold. Br. J. Ophthalmol., 95 (3), 410-414.
Funamoto, S., Nam, K., Kimura, T., Murakoshi, A., Hashimoto, Y., Niwaya K, ..., & Kishida, A. (2010). The use of high-hydrostatic pressure treatment to decellularize blood vessels. Biomaterials, 31 (13), 3590-3595.
Riau, A.K., Beuerman, R.W., Lim, L.S., & Mehta, J.S. (2010). Preservation, sterilization and de-epithelialization of human amniotic membrane for use in ocular surface reconstruction. Biomaterials, 31 (2), 216-225.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2021 Achievements of Clinical and Experimental Medicine
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
