Construction of periodontal biomimetic membrane with
electrospun poly(lactic-co-glycolic acid) nanofibers and electrosprayed
chitosan microspheres

doi:10.3969/j.issn.2095-4344.2212

Abstract

Abstract:

BACKGROUND: When the teeth are separated from the alveolar fossa, the periodontal membrane breaks, and the residual periodontal membrane on the avulsed tooth root surface changes from three-dimensional to two-dimensional, thus losing the role of scaffold, and leading to root bone adhesion after replantation of avulsed tooth. How to develop a three-dimensional sustained-release scaffold material that can adhere to the root surface with a certain thickness and strength is one of the key factors for successful regeneration of avulsed tooth periodontal membrane.

OBJECTIVE: To construct a three-dimensional periodontal biomimetic membrane that can adhere to the avulsed tooth root surface and allow sustained-release of growth factors.

METHODS: Poly(lactic-co-glycolic acid) (PLGA) membrane was prepared using electrospinning technique. The effects of dichloromethane and dimethylformamide mixture, hexafluoroisopropanol, and trichloromethane on electrospun membrane were investigated to obtain the optimal electrospinning solvent. Chitosan microspheres were prepared by electrospray and ion cross-linking techniques. The effects of molecular weight (50,000,100,000) and mass concentration (10, 20 g/L) of chitosan, sodium tripolyphosphate concentration (2%, 5%, 10%) and voltage (14, 28 kV) on chitosan microspheres were studied to screen the optimum parameters. Chitosan microspheres containing stromal cell-derived factor-1 (optimal parameter design) were constructed. The release rate of stromal cell-derived factor-1 alpha in vitro was determined. First, the root surface of teeth was wrapped with electrospun PLGA membrane, then chitosan microspheres were dripped on the surface, and finally the surface was wrapped with a thin layer of electrospun PLGA. Thus, PLGA-chitosan-PLGA biomimetic membrane was constructed.

RESULTS AND CONCLUSION: Electrospun PLGA membrane prepared with hexafluoroisopropanol as electrospinning solvent had the smallest average diameter and the largest porosity. When the relative molecular weight of chitosan was 50,000 and the mass concentration was 20 g/L, the size of chitosan microspheres was basically the same, and the average diameter was 366.6 μm. In addition, chitosan microsphere had good monodispersity, fullness, and stability. Chitosan microspheres formed under 28 kV voltage and were more in line with the requirements of biomimetic membrane for avulsed tooth. The surface of microspheres prepared by 5% sodium tripolyphosphate had medium-sized pores, which are most conducive to clinical periodontal membrane regeneration. Chitosan microspheres can sustainably release stromal cell derived factor 1alpha for about 1 month. In this study, we constructed a three-dimensional PLGA-chitosan-PLGA periodontal biomimetic membrane that can adhere to the avulsed tooth root surface and allow sustained-release of growth factors and obtained the optimal parameters of constructing the periodontal biomimetic membrane. Based on the PLGA-chitosan-PLGA periodontal biomimetic membrane, the effect and mechanism of tissue engineering on replantation of avulsed tooth can be further studied.

Key words: avulsed tooth, periodontal regeneration, biomimetic membrane, PLGA, chitosan microspheres, controlled-release, electrospinning, electrospray

CLC Number:

Feng Xiaoxia, Hou Weiwei, Jin Xiaoting, Wang Xinhua. Construction of periodontal biomimetic membrane with electrospun poly(lactic-co-glycolic acid) nanofibers and electrosprayed chitosan microspheres[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(4): 511-516.

Figures/Tables 3

2.2 不同参数对电喷壳聚糖微球直径及形貌的影响 2.2.1 壳聚糖质量浓度及相对分子质量对微球的影响光镜下，相对分子质量5万、质量浓度10 g/L壳聚糖形成的微球比较散，大小不一，平均直径为362.1 μm，易破；相对分子质量5万、质量浓度20 g/L壳聚糖形成的微球大小基本一致，平均直径366.6 μm，单分散性好、饱满、稳定；相对分子质量10万、质量浓度10 g/L壳聚糖形成的微球比较大，最大直径可达510.88 μm，不稳定，见图2A，2C-a。因此，选定相对分子质量5万、质量浓度20 g/L的壳聚糖进行下一步实验。 2.2.2 电压对电微球直径的影响 14 kV低压下，壳聚糖微球的平均直径366.6 μm；28 kV高压下，壳聚糖微球的平均直径在112.1 μm，两者之间比较差异有非常显著性意义(P < 0.000 1)，见图2C-b，此结果提示电喷技术中电压参数是影响壳聚糖微球直径大小的重要因素之一。随着电压升高，壳聚糖微球的直径变小，高压下电喷微球直径在100 μm，牙周膜宽度100-380 μm，直径100 μm微球便于用于仿生牙周膜支架，提示高压下形成的壳聚糖微球更符合脱位牙仿生膜的要求。 2.2.3 三聚磷酸钠接受液浓度对微球微观结构的影响如图2B所示，扫描电镜示微球多孔结构，表面光滑，均一性好，无粘连。不同浓度三聚磷酸钠制备的壳聚糖微球大小一致，但其微观结构形貌孔径差别很大，3组微球微观结构孔径比较差异有显著性意义(P < 0.05)，见图2C-c。 2%三聚磷酸钠所得的壳聚糖表面微观结构孔径最小，平均值为0.915 7 μm；5%三聚磷酸钠所得的壳聚糖表面微观结构孔径居中，平均值为4.775 μm；10%三聚磷酸钠所得的壳聚糖孔径最大，平均直径为9.407 μm。可见接受液三聚磷酸钠浓度明显影响壳聚糖微球表观结构孔径的大小。壳聚糖微观孔径太大有可能影响细胞的黏附，孔径太小有可能影响细胞的穿入，所以5%三聚磷酸钠下壳聚糖微球微观结构可能是最有利于临床牙周膜再生的支架结构。 2.2.4 含基质细胞衍生因子1α壳聚糖微球的缓释速率每样本含有200 ng基质细胞衍生因子1α，37 ℃ 54 r/min恒温摇床振荡24 h后，生长因子释放速率达5 μg/L，此后释放率继续上升，4 d达最大，近10 μg/L，但之后缓慢下降，于25 d释放速率减至0.5 μg/L，见图2D。总之，壳聚糖微球具有大约1个月的缓释效能。 "

References

[1] GLENDOR U.Epidemiology of traumatic dental injuries--a 12 year review of the literature. Dent Traumatol.2008;24(6):603-611.
[2] PRIYA MH, TAMBAKAD PB, NAIDU J.Pulp and Periodontal Regeneration of an Avulsed Permanent Mature Incisor Using Platelet-rich Plasma after Delayed Replantation: A 12-month Clinical Case Study.J Endod.2016;42(1):66-71.
[3] CHEW JRJ, CHUAH SJ, TEO KYW, et al.Mesenchymal stem cell exosomes enhance periodontal ligament cell functions and promote periodontal regeneration. Acta Biomaterialia.2019;89:252-264.
[4] DEMIREL S, YALVAC ME, TAPSIN S, et al.Tooth replantation with adipose tissue stem cells and fibrin sealant: microscopic analysis of rat’s teeth.Springerplus.2016;l5(1):656.
[5] VENKATAIAH VS, HANDA K, NJUGUNA MM, et al.Periodontal Regeneration by Allogeneic Transplantation of Adipose Tissue Derived Multi-Lineage Progenitor Stem Cells in vivo. Sci Rep.2019;9(1):921.
[6] ZHU W, ZHANG Q, ZHANG Y, et al.PDL regeneration via cell homing in delayed replantation of avulsed teeth.J Transl Med.2015;13(1):357.
[7] TABATA Y, YAMADA K, MIYAMOTO S, et al.Bone regeneration by basic fibroblast growth factor complexed with biodegradable hydrogels. Biomaterials.1998;19(7-9):807-815.
[8] ASPENBERG P, ALBREKTSSON T, THORNGREN KG. Local application of growth-factor IGF-1 to healing bone. Experiments with a titanium chamber in rabbits.Acta Orthop Scand.1989;60(5):607-610.
[9] SINPREECHANON P, BOONZONG U, SRICHOLPECH M. Comparative evaluation of periodontal ligament fibroblasts stored in different types of milk: effects on viability and biosynthesis of collagen. Eur J Oral Sci. 2019;127(4):323-332.
[10] PHAM QP, SHARMA U, MIKOS AG.Electrospinning of Polymeric Nanofibers for Tissue Engineering Applications: A Review.Tissue Eng. 2006;12(5):1197-1211.
[11] JIANG W, LI L, ZHANG D, et al.Incorporation of aligned PCL-PEG nanofibers into porous chitosan scaffolds improved the orientation of collagen fibers in regenerated periodontium. Acta Biomater. 2015;25: 240-252.
[12] CHEN G, CHEN J, YANG B, et al.Combination of aligned PLGA/Gelatin electrospun sheets, native dental pulp extracellular matrix and treated dentin matrix as substrates for tooth root regeneration. Biomaterials. 2015;52:56-70.
[13] DAI T, TANAKA M, HUANG YY, et al.Chitosan preparations for wounds and burns: antimicrobial and wound-healing effects.Expert Rev Anti Infect Ther.2011;9(7):857-879.
[14] ATAY E, FABRA MJ, MARTÍNEZ-SANZ M, et al.Development and characterization of chitosan/gelatin electrosprayed microparticles as food grade delivery vehicles for anthocyanin extracts.Food Hydrocoll. 2018;77:699-710.
[15] JU X, WANG X, LIU Z, et al.Red-blood-cell-shaped chitosan microparticles prepared by electrospraying.Particuology. 2017;30: 151-157.
[16] DASH M,CHIELLINI F,OTTENBRITE RM,et al.Chitosan—A versatile semi-synthetic polymer in biomedical applications.Prog Polym Sci. 2011;36(8):981-1014.
[17] HE Y, JIN Y, WANG X, et al.An Antimicrobial Peptide-Loaded Gelatin/Chitosan Nanofibrous Membrane Fabricated by Sequential Layer-by-Layer Electrospinning and Electrospraying Techniques. Nanomaterials (Basel).2018;8(5). pii: E327. doi: 10.3390/nano8050327.
[18] KAPOOR DN, BHATIA A, KAUR R, et al.PLGA: a unique polymer for drug delivery.Ther Deliv.2015;6(1):41-58.
[19] MARQUEZ-CURTIS LA, JANOWSKA-WIECZOREK A.Enhancing the migration ability of mesenchymal stromal cells by targeting the SDF-1/CXCR4 axis.Biomed Res Int.2013;2013:561098.
[20] FUJIO M, YAMAMOTO A, ANDO Y, et al.Stromal cell-derived factor-1 enhances distraction osteogenesis-mediated skeletal tissue regeneration through the recruitment of endothelial precursors.Bone. 2011;49(4):693-700.
[21] KIM K, LEE CH, KIM BK, et al.Anatomically shaped tooth and periodontal regeneration by cell homing.J Dent Res. 2010;89(8):842-847.
[22] BODMEIER R, CHEN HG,PAERATAKUL O.A novel approach to the oral delivery of micro- or nanoparticles.Pharm Res.1989;6(5):413-417.
[23] SHU XZ, ZHU KJ.Controlled drug release properties of ionically cross-linked chitosan beads: the influence of anion structure.Int J Pharm.2002;233(1):217-225.
[24] 周旋.壳聚糖微球制备优化及其乙酰化微球作为潜在栓塞材料的研究[D].青岛:中国海洋大学,2012.
[25] HASSANI S, LAOUINI A, Fessi H, et al.Preparation of chitosan–TPP nanoparticles using microengineered membranes – Effect of parameters and encapsulation of tacrine.Colloid and Surface A. 2015;482:34-43.
[26] XU Y, HANNA MA.Electrosprayed bovine serum albumin-loaded tripolyphosphate cross-linked chitosan capsules: synthesis and characterization.J Microencapsul.2007;24(2):143-151.
[27] WANG XX, JU XJ, SUN SX, et al.Monodisperse erythrocyte-sized and acid-soluble chitosan microspheres prepared via electrospraying.RSC Adv.2015;5(43):34243-34250.
[28] MORENO JAS, MENDES AC, Stephansen K, et al.Development of electrosprayed mucoadhesive chitosan microparticles.Carbohydr Polym.2018;190:240-247.