Preliminary study of silk fibroin porous scaffold for oral soft-tissue thickening

doi:10.7518/hxkq.2022.05.003

Abstract

Abstract:

Objective This study aimed to investigate the feasibility of three different concentrations of silk-fibroin porous scaffolds applied to oral soft-tissue thickening in vivo. Methods Silk-fibroin scaffolds with three different concentrations (1 wt%, 3 wt%, and 5 wt%; denoted as SF1, SF3, and SF5, respectively) were prepared by freeze drying and methanol enhancement. The scaffolds were characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), and thermogravimetric analysis. Pore size, porosity, and in vitro degradation rate were also evaluated. The three groups of scaffold materials (the experimental sides) and the collagen matrix (the control side) were implanted into the oral mucosa of New Zealand white rabbits. Changed in mucosa thickness before and 3 months after operation were compared. The in vivo metabolism and regeneration effect of each group were observed by histological hematoxylin-eosin (HE) and Masson staining. Results SEM showed that the three groups of scaffolds were all cross-linked porous structures. XRD and FTIR showed that the three scaffolds were dominated by a relatively stable Silk Ⅱ structure, which degraded more slowly in vitro. Among them, SF3 had the largest pore size (133.40 μm±22.85 μm) and moderate porosity (90.05%±6.68%). In vivo results showed that the thickening effect of SF1 was similar to that of the control group because of insufficient space-maintenance property. Meanwhile, the properties of SF3 and SF5 were more stable, and the thickening effect was significantly better than those of the control group. However, unlike SF5 that induced obvious inflammation, SF3 showed better degradation, more fibrosis and angiogenesis, and less inflammatory response in vivo. Conclusion Silk-fibroin scaffolds can be applied to effectively thicken soft tissues, among which SF3 (3 wt%) silk fibroin scaffold exhibited the best physicochemical properties, histocompatibility, and mucosal-thickening effect.

Key words: scaffold, silk fibroin, soft tissue thickening, animal experiment

CLC Number:

R 782.2

Li Dexiong, Cao Runyuan, Chen Jiang. Preliminary study of silk fibroin porous scaffold for oral soft-tissue thickening[J]. West China Journal of Stomatology, 2022, 40(5): 513-521.

Figures/Tables 6

Fig 1

Fig 2

Fig 3

Fig 4

Fig 5

Fig 6

References 36

1	Vallecillo C, Toledano-Osorio M, Vallecillo-Rivas M, et al. Collagen matrix vs. autogenous connective tissue graft for soft tissue augmentation: a systematic review and meta-analysis[J]. Polymers (Basel), 2021, 13(11): 1810.
2	孟逍逸, 薛毅. 前牙美学区种植的软组织增量研究进展[J]. 口腔颌面修复学杂志, 2018, 19(1): 59-64.
	Meng XY, Xue Y. Soft tissue augmentation of dental implant in esthetic zone of anterior teeth[J]. Chin J Prosthodont, 2018, 19(1): 59-64.
3	Puzio M, Hadzik J, Błaszczyszyn A, et al. Soft tissue augmentation around dental implants with connective tissue graft (CTG) and xenogenic collagen matrix (XC-M). 1-year randomized control trail[J]. Ann Anat, 2020, 230: 151484.
4	Bakkali S, Rizo-Gorrita M, Romero-Ruiz MM, et al. Efficacy of different surgical techniques for peri-implant tissue preservation in immediate implant placement: a systematic review and meta-analysis[J]. Clin Oral Invest, 2021, 25(4):1655-1675.
5	Panwar M, Kosala M, Malik D, et al. Comparison of acellular dermal matrix allografts and connective tissue autografts in soft-tissue augmentation around immediate implants: a pilot study[J]. Med J Armed Forces India, 2021(7): 029.
6	Kroiss S, Rathe F, Sader R, et al. Acellular dermal matrix allograft versus autogenous connective tissue grafts for thickening soft tissue and covering multiple gingival recessions: a 5-year preference clinical study[J]. Quintessence Int, 2019, 50(4): 278-285.
7	Cairo F, Barbato L, Tonelli P, et al. Xenogeneic collagen matrix versus connective tissue graft for buccal soft tissue augmentation at implant site. A randomized, controlled clinical trial[J]. J Clin Periodontol, 2017, 44(7): 769-776.
8	Puzio M, Błaszczyszyn A, Hadzik J, et al. Ultrasound assessment of soft tissue augmentation around implants in the aesthetic zone using a connective tissue graft and xenogeneic collagen matrix-1-year randomised follow-up[J]. Ann Anat, 2018, 217: 129-141.
9	Rothamel D, Benner M, Fienitz T, et al. Biodegradation pattern and tissue integration of native and cross-linked porcine collagen soft tissue augmentation matrices—an experimental study in the rat[J]. Head Face Med, 2014, 10: 10.
10	Rothamel D, Schwarz F, Sager M, et al. Biodegradation of differently cross-linked collagen membranes: an experimental study in the rat[J]. Clin Oral Implants Res, 2005, 16(3): 369-378.
11	Melke J, Midha S, Ghosh S, et al. Silk fibroin as biomaterial for bone tissue engineering[J]. Acta Biomater, 2016, 31: 1-16.
12	Holland C, Numata K, Rnjak-Kovacina J, et al. The biomedical use of silk: past, present, future[J]. Adv Healthc Mater, 2019, 8(1): e1800465.
13	Zhang Q, Zhao Y, Yan S, et al. Preparation of uniaxial multichannel silk fibroin scaffolds for guiding primary neurons[J]. Acta Biomater, 2012, 8(7): 2628-2638.
14	Huang W, Ling S, Li C, et al. Silkworm silk-based materials and devices generated using bio-nanotechnology[J]. Chem Soc Rev, 2018, 47(17): 6486-6504.
15	Li X, Yan S, Qu J, et al. Soft freezing-induced self-assembly of silk fibroin for tunable gelation[J]. Int J Biol Macromol, 2018, 117: 691-695.
16	Cheng G, Davoudi Z, Xing X, et al. Advanced silk fibroin biomaterials for cartilage regeneration[J]. ACS Biomater Sci Eng, 2018, 4(8): 2704-2715.
17	Wang XL, Guo CC, Guo LN, et al. Radially aligned porous silk fibroin scaffolds as functional templates for engineering human biomimetic hepatic lobules[J]. ACS Appl Mater Interfaces, 2022, 14(1): 201-213.
18	Kim UJ, Park J, Kim HJ, et al. Three-dimensional aqueous-derived biomaterial scaffolds from silk fibroin[J]. Biomaterials, 2005, 26(15): 2775-2785.
19	Nazarov R, Jin HJ, Kaplan DL. Porous 3-D scaffolds from regenerated silk fibroin[J]. Biomacromolecules, 2004, 5(3): 718-726.
20	Lu Q, Wang X, Lu S, et al. Nanofibrous architecture of silk fibroin scaffolds prepared with a mild self-assembly process[J]. Biomaterials, 2011, 32(4): 1059-1067.
21	Yao D, Qian Z, Zhou J, et al. Facile incorporation of REDV into porous silk fibroin scaffolds for enhancing vascularization of thick tissues[J]. Mater Sci Eng C Mater Biol Appl, 2018, 93: 96-105.
22	Han H, Ning H, Liu S, et al. Silk biomaterials with vascularization capacity[J]. Adv Funct Mater, 2016, 26(3): 421-436.
23	Sang Y, Li M, Liu J, et al. Biomimetic silk scaffolds with an amorphous structure for soft tissue engineering[J]. ACS Appl Mater Interfaces, 2018, 10(11): 9290-9300.
24	Berry CC, Campbell G, Spadiccino A, et al. The influence of microscale topography on fibroblast attachment and motility[J]. Biomaterials, 2004, 25(26): 5781-5788.
25	Salem AK, Stevens R, Pearson RG, et al. Interactions of 3T3 fibroblasts and endothelial cells with defined pore features[J]. J Biomed Mater Res, 2002, 61(2): 212-217.
26	Ho MH, Kuo PY, Hsieh HJ, et al. Preparation of porous scaffolds by using freeze-extraction and freeze-gelation methods[J]. Biomaterials, 2004, 25(1): 129-138.
27	Zeltinger J, Sherwood JK, Graham DA, et al. Effect of pore size and void fraction on cellular adhesion, proliferation, and matrix deposition[J]. Tissue Eng, 2001, 7(5): 557-572.
28	Cui L, Li J, Long Y, et al. Vascularization of LBL structured nanofibrous matrices with endothelial cells for tissue regeneration[J]. RSC Adv, 2017, 7(19): 11462-11477.
29	Han H, Ning H, Liu S, et al. Silk biomaterials with vascularization capacity[J]. Adv Funct Mater, 2016, 26(3): 421-436.
30	Bhardwaj N, Chakraborty S, Kundu SC. Freeze-gelled silk fibroin protein scaffolds for potential applications in soft tissue engineering[J]. Int J Biol Macromol, 2011, 49(3): 260-267.
31	Etienne O, Schneider A, Kluge JA, et al. Soft tissue augmentation using silk gels: an in vitro and in vivo study[J]. J Periodontol, 2009, 80(11): 1852-1858.
32	Jansen RG, van Kuppevelt TH, Daamen WF, et al. Tissue reactions to collagen scaffolds in the oral mucosa and skin of rats: environmental and mechanical factors[J]. Arch Oral Biol, 2008, 53(4): 376-387.
33	Schenk RK, Buser D, Hardwick WR, et al. Healing pattern of bone regeneration in membrane-protected defects: a histologic study in the canine mandible[J]. Int J Oral Maxillofac Implants, 1994, 9(1): 13-29.
34	Lundgren AK, Sennerby L, Lundgren D. Guided jaw-bone regeneration using an experimental rabbit model[J]. Int J Oral Maxillofac Surg, 1998, 27(2): 135-140.
35	Mathes SH, Wohlwend L, Uebersax L, et al. A bioreactor test system to mimic the biological and mechanical environment of oral soft tissues and to evaluate substitutes for connective tissue grafts[J]. Biotechnol Bioeng, 2010, 107(6): 1029-1039.
36	Marzadori M, Stefanini M, Mazzotti C, et al. Soft-tissue augmentation procedures in edentulous esthetic areas[J]. Periodontol 2000, 2018, 77(1): 111-122.

[1]	Liu Yiping, Wang Jue, Tian Zilu, Zhai Peisong, Wang Zhanqi, Zhou Yanmin, Ni Shilei. Effects of scaffold microstructure and mechanical properties on regeneration of tubular dentin [J]. West China Journal of Stomatology, 2020, 38(3): 314-318.
[2]	Xinxin Ding, Yanmin Zhou, Xing-chen Xiang, Lin Meng, Qin Qin, Shan Ye. Research progress on chitosan composite scaffolds in bone tissue engineering [J]. West China Journal of Stomatology, 2018, 36(4): 441-446.
[3]	Binhong Teng, Yanhong Zhao, Lianyong Wang, Qiang Yang, Hongfa Li, Yunjie Li. Preparation and characterization of oriented scaffolds derived from cartilage extracellular matrix and silk fibroin [J]. West China Journal of Stomatology, 2018, 36(1): 17-22.
[4]	Kun Li, Yanhong Zhao, Chen Xu, Lianyong Wang, Qiang Yang, Hongfa Li, Binhong Teng. Development and characterization of oriented scaffolds derived from cartilage extracellular matrix [J]. West China Journal of Stomatology, 2017, 35(1): 51-56.
[5]	Xi Weihong, Wang Zhen, Zhu Hongshui, Li Xiaofeng, Xiong Yuanfei. Synthesis and characteristics of integrated bionic mandibular condylar scaffold [J]. West China Journal of Stomatology, 2016, 34(1): 68-72.
[6]	Zhang Yan, Jiang Xinquan, Zhang Xiuli, Wang Deping, Zhen Lei.. Cytocompatibility of two porous bioactive glass-ceramic in vitro [J]. West China Journal of Stomatology, 2013, 31(3): 294-299.
[7]	Zhang Kefu1, Zhang Shu1, Luo Zhiqiang1, Wang Jing1，2, Wang Tao3, Ou Guomin1，4, Wang Hu1，5. Biocompatibility of porous calcium phosphate ceramic nanocomposite [J]. West China Journal of Stomatology, 2012, 30(2): 209-213.
[8]	LAI Ren-fa, ZHAO Qing-tong, LIU Xiang-ning, SHEN Shan. Study on osteogenic ability of chitosan/β-tricalcium phosphate scaffold combined with human bone morphogenetic protein [J]. West China Journal of Stomatology, 2010, 28(05): 464-467.
[9]	WANG Lei1, LI Yao -jun2, ZHANG Yan3, PAN Ke -feng3, HUANG Yuan -liang2, LIU Chang -sheng4, JIANG Xin-quan5. The experimental study on porous calcium phosphate cement with bone marrow stromal cells for bone tissue engineering [J]. West China Journal of Stomatology, 2010, 28(03): 315-318.
[10]	LI Yi1, RAN Wei2, WANG Gai-ling1, JING Xiang-dong1. Biocompatibility of new bone tissue engineering scaffolds in vivo [J]. West China Journal of Stomatology, 2009, 27(04): 447-450.
[11]	WANG Lixia1，2, ZHAO Huan1, JIANG Bo3, DING Yi1. Adhesion and growth of human periodontal ligament cells on hyaluronic acid/collagen scaffold [J]. West China Journal of Stomatology, 2009, 27(02): 220-223.
[12]	LIU Shao-hua1, WEI Feng-cai1, ZHANG Dong1, SUN Shan-zhen2, ZHAO Hua-qiang2, LI Guo-ju2. Experimental Study of Mandibular Periosteal Distraction in Rabbits [J]. West China Journal of Stomatology, 2006, 24(03): 273-275.
[13]	NONG Xiao-lin1, WANG Da-zhang2,MENGMin1,ZHOUNuo1,MENGNing1,LIJia-quan3,ZHANGHong4. Combining of TNP-470 and 5-Fu in Inhibition of Adenoid Cystic Carcinoma in Nude Mice Model [J]. West China Journal of Stomatology, 2004, 22(04): 267-270.