E V Isaeva, A A Kisel, E E Beketov, G A Demyashkin, N D Yakovleva, T S Lagoda, N V Arguchinskaya, D S Baranovsky, S A Ivanov, P V Shegay, A D Kaprin
{"title":"胶原蛋白和 GelMA 对在体内支架中保留肋软骨细胞表型的影响","authors":"E V Isaeva, A A Kisel, E E Beketov, G A Demyashkin, N D Yakovleva, T S Lagoda, N V Arguchinskaya, D S Baranovsky, S A Ivanov, P V Shegay, A D Kaprin","doi":"10.17691/stm2023.15.2.01","DOIUrl":null,"url":null,"abstract":"<p><p><b>The aim of the study</b> was to compare type I collagen-based and methacryloyl gelatin-based (GelMA) hydrogels by their ability to form hyaline cartilage in animals after subcutaneous implantation of scaffolds.</p><p><strong>Materials and methods: </strong>Chondrocytes were isolated from the costal cartilage of newborn rats using 0.15% collagenase solution in DMEM. The cells was characterized by glycosaminoglycan staining with alcian blue. Chondrocyte scaffolds were obtained from 4% type I porcine atelocollagen and 10% GelMA by micromolding and then implanted subcutaneously into the withers of two groups of Wistar rats. Histological and immunohistochemical studies were performed on days 12 and 26 after implantation. Tissue samples were stained with hematoxylin and eosin, alcian blue; type I and type II collagens were identified by the corresponding antibodies.</p><p><strong>Results: </strong>The implanted scaffolds induced a moderate inflammatory response in both groups when implanted in animals. By day 26 after implantation, both collagen and GelMA had almost completely resorbed. Cartilage tissue formation was observed in both animal groups. The newly formed tissue was stained intensively with alcian blue, and the cells were positive for both types of collagen. Cartilage tissue was formed among muscle fibers.</p><p><strong>Conclusion: </strong>The ability of collagen type I and GelMA hydrogels to form hyaline cartilage in animals after subcutaneous implantation of scaffolds was studied. Both collagen and GelMA contributed to formation of hyaline-like cartilage tissue type in animals, but the chondrocyte phenotype is characterized as mixed. Additional detailed studies of possible mechanisms of chondrogenesis under the influence of each of the hydrogels are needed.</p>","PeriodicalId":51886,"journal":{"name":"Sovremennye Tehnologii v Medicine","volume":null,"pages":null},"PeriodicalIF":1.1000,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10306965/pdf/","citationCount":"0","resultStr":"{\"title\":\"Effect of Collagen and GelMA on Preservation of the Costal Chondrocytes' Phenotype in a Scaffold <i>in vivo</i>.\",\"authors\":\"E V Isaeva, A A Kisel, E E Beketov, G A Demyashkin, N D Yakovleva, T S Lagoda, N V Arguchinskaya, D S Baranovsky, S A Ivanov, P V Shegay, A D Kaprin\",\"doi\":\"10.17691/stm2023.15.2.01\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><b>The aim of the study</b> was to compare type I collagen-based and methacryloyl gelatin-based (GelMA) hydrogels by their ability to form hyaline cartilage in animals after subcutaneous implantation of scaffolds.</p><p><strong>Materials and methods: </strong>Chondrocytes were isolated from the costal cartilage of newborn rats using 0.15% collagenase solution in DMEM. The cells was characterized by glycosaminoglycan staining with alcian blue. Chondrocyte scaffolds were obtained from 4% type I porcine atelocollagen and 10% GelMA by micromolding and then implanted subcutaneously into the withers of two groups of Wistar rats. Histological and immunohistochemical studies were performed on days 12 and 26 after implantation. Tissue samples were stained with hematoxylin and eosin, alcian blue; type I and type II collagens were identified by the corresponding antibodies.</p><p><strong>Results: </strong>The implanted scaffolds induced a moderate inflammatory response in both groups when implanted in animals. By day 26 after implantation, both collagen and GelMA had almost completely resorbed. Cartilage tissue formation was observed in both animal groups. The newly formed tissue was stained intensively with alcian blue, and the cells were positive for both types of collagen. Cartilage tissue was formed among muscle fibers.</p><p><strong>Conclusion: </strong>The ability of collagen type I and GelMA hydrogels to form hyaline cartilage in animals after subcutaneous implantation of scaffolds was studied. Both collagen and GelMA contributed to formation of hyaline-like cartilage tissue type in animals, but the chondrocyte phenotype is characterized as mixed. Additional detailed studies of possible mechanisms of chondrogenesis under the influence of each of the hydrogels are needed.</p>\",\"PeriodicalId\":51886,\"journal\":{\"name\":\"Sovremennye Tehnologii v Medicine\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.1000,\"publicationDate\":\"2023-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10306965/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Sovremennye Tehnologii v Medicine\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.17691/stm2023.15.2.01\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2023/3/29 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q4\",\"JCRName\":\"MEDICINE, RESEARCH & EXPERIMENTAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Sovremennye Tehnologii v Medicine","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.17691/stm2023.15.2.01","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2023/3/29 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"MEDICINE, RESEARCH & EXPERIMENTAL","Score":null,"Total":0}
引用次数: 0
摘要
该研究旨在比较 I 型胶原水凝胶和甲基丙烯酰明胶(GelMA)水凝胶在动物皮下植入支架后形成透明软骨的能力:使用 DMEM 中的 0.15% 胶原酶溶液从新生大鼠肋软骨中分离软骨细胞。用乙酰胆碱对细胞进行糖胺聚糖染色。软骨细胞支架由 4% I 型猪鹅胶原和 10% GelMA 通过微成型制成,然后植入两组 Wistar 大鼠的腰部皮下。植入后第 12 天和第 26 天进行组织学和免疫组化研究。组织样本用苏木精、伊红和藻蓝染色,用相应的抗体鉴定 I 型和 II 型胶原:结果:两组动物植入支架后都出现了中度炎症反应。植入后第 26 天,胶原蛋白和 GelMA 几乎完全吸收。两组动物都观察到了软骨组织的形成。新形成的组织用阿尔金山蓝染色,细胞对两种胶原均呈阳性反应。软骨组织在肌肉纤维间形成:研究了 I 型胶原蛋白和 GelMA 水凝胶在皮下植入支架后在动物体内形成透明软骨的能力。胶原蛋白和 GelMA 都有助于在动物体内形成透明软骨组织类型,但软骨细胞表型的特点是混合型。还需要对每种水凝胶影响下软骨生成的可能机制进行更详细的研究。
Effect of Collagen and GelMA on Preservation of the Costal Chondrocytes' Phenotype in a Scaffold in vivo.
The aim of the study was to compare type I collagen-based and methacryloyl gelatin-based (GelMA) hydrogels by their ability to form hyaline cartilage in animals after subcutaneous implantation of scaffolds.
Materials and methods: Chondrocytes were isolated from the costal cartilage of newborn rats using 0.15% collagenase solution in DMEM. The cells was characterized by glycosaminoglycan staining with alcian blue. Chondrocyte scaffolds were obtained from 4% type I porcine atelocollagen and 10% GelMA by micromolding and then implanted subcutaneously into the withers of two groups of Wistar rats. Histological and immunohistochemical studies were performed on days 12 and 26 after implantation. Tissue samples were stained with hematoxylin and eosin, alcian blue; type I and type II collagens were identified by the corresponding antibodies.
Results: The implanted scaffolds induced a moderate inflammatory response in both groups when implanted in animals. By day 26 after implantation, both collagen and GelMA had almost completely resorbed. Cartilage tissue formation was observed in both animal groups. The newly formed tissue was stained intensively with alcian blue, and the cells were positive for both types of collagen. Cartilage tissue was formed among muscle fibers.
Conclusion: The ability of collagen type I and GelMA hydrogels to form hyaline cartilage in animals after subcutaneous implantation of scaffolds was studied. Both collagen and GelMA contributed to formation of hyaline-like cartilage tissue type in animals, but the chondrocyte phenotype is characterized as mixed. Additional detailed studies of possible mechanisms of chondrogenesis under the influence of each of the hydrogels are needed.