In vitro morphological observation on the effect of exogenous collagen on the construction of tissue-engineered tendon
- VernacularTitle:外源性胶原对组织工程肌腱构建影响的体外形态学观察
- Author:
Yue FANG
;
Zhou XIANG
;
Zhiming YANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2006;10(29):169-172
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Collagen type Ⅰ possesses low antigenicity and provides special functions of scaffold structure for cell migration, proliferation and secretion.OBJECTIVE: To observe the morphological change after combination of transformed human embryonic tendon cells and artificial materials with collagen type Ⅰ, so as to provide the experimental basis for constructing tissue-engineered muscle tendon in vitro.DESIGN: Randomized and controlled observation.SETTING: Department of Orthopaedics, West China Hospital, Sichuan University.MATERIALS: This experiment was carried out at the Department of Transplantation Immunology, West China University of Medical Sciences from July to September 1998. Human hair, carbon fiber and polyglycolic acid were combined with collagen and then set corresponding groups respectively. Simply hair, carbon fiber and polyglycolic acid control groups were set respectively. Artificial materials: Healthy adult hair (30 cm in length, 80 g in weight), carbon fiber knitted strip (Lanzhou carbon factory )and polyglyolic acid fasciculi (Johnson , American) were used in this experiment.METHODS: ①In the group of human hair combined with collagen,nine healthy adult hairs were chosen . Three hairs as one bundle were knitted into tendon with 0.8 cm in length and 0.4 cm in diameter and a knot was tied respectively at the two ends. ②In the carbon fiber combined with collagen group, the purchased carbon fiber knitted strip was separated and made into artificial materials with the same diameter and length as those of human hair tendon. Two ends of the artificial materials were packed with 5/0 silk thread. ③In the polyglycolic acid combined with collagen group, three polyglycolic acid fasciculi were weaved into bundle with the same diameter and length as those of human tendon,and two ends were packed with 5/0 silk thread. ④The process of knitting scaffold materials was the same in control groups as in the corresponding experimental groups. ⑤ Five pieces were prepared in each scaffold material. After sterilized by 60Co, the scaffold materials were put in collagen type Ⅰ solution for 0.5 hour, then taken out and dried at roomtemperature for use in the experimental groups. ⑥transformed human embryonic tendon cell, density for 3×106/mm3, was used for combined culture in each group, and cell number and morphology were observed at different time points after culture under an inverted microscope and scanning electron microscope.MAIN OUTCOME MEASURES: ① Observation of cell growth and adherence under an inverted microscope ② Observation of cell morphology and material adherence under scanning electron microscope RESULTS: ① Observation of cell growth and adherenceunder an inverted microscope :In each experimental group, at two hours after culture, cells gathered around the materials, and some cells adhered to materials presented round; At three days after culture, the number of cells adhered to materials began to increase, morphology changed from round to oval, and the co-existence of round and oval cells was seen;While in each control group, at two and three hours after culture, few cells adhered to the three materials. On day five after culture, in each experimental group, cell morphology turned into fusiform, the number of cells increased further and a great many fusiform cells grew among carbon fiber materials; while in the control group, most cells presented round and ellipse, and the number of cells was less as compared with the corresponding experimental group. ② Cell morphology and material adherence under scanning electron microscope: On day 5 after in vitro culture, most tendon cells on the surface of materials presented spindle-shape in each experimental group. Apophysis stretched out from the tendon cells, grew along longitudinal axon of the materials. Silk and piece-like substance appeared among the apophysis and weaved each other. While there were fewer cells adhered in control group than in corresponding experimental group, cells also presented fusiform or round, but no silk or piece-like substance was found, also no apophysis appeared.CONCLUSION: Human hair, carbon fiber and polyglycolic acid combined with collagen type Ⅰ will facilitate cell adherence. Cells increase around the materials and co-existence of round and oval cells could be seen, and cells turned into fusiform, suggesting that exogenous collagen facilitate the cells to adhere onto the materials and proliferate.
