PRELIMINARY STUDY ON IN VITRO TENDON ENGINEE-RING USING TENOCYTES AND POLYGLYCOLIC ACIDS

Objective To investigate the feasibility of tendon engineering in vitro using tenocyws and polyglycolic acids （ PGA ）. Methods Tenocytes were isolated by tissue explant method and expanded in vitro. Cells of the second passage were collected and seeded onto PGA scaffolds made from PGA unwoven fibers at the density of 20 × 10^6 cells/ml. At 1 week postseeding ,the constructs were divided into three groups as follows： cell-scaffold constructs under constant tension generated by a U-shaped spring as the experimental group （ n = 5 ）, cell-scaffold constructs under no tension as control group 1 （ n = 4 ）, cell-free scaffolds under constant tension as control group 2 （n =3）. Samples were harvested at 2, 4 and 6 weeks for histological and immunohistochemical （ IHC ） examinations. Transmission electron microscopy （TEM） and mechanical test were performed to evaluate the constructs of 6 weeks. Results At 2 weeks, the constructs were mainly composed of undegraded PGA fibers. Gross and histological examination revealed no difference between the groups. At 4 weeks, neo-tendon was visible through gross observation in experimental group and control group 1. Histology and immunohistochemistry revealed the formation of collagen fibers. While in control group 2, PGA fibers were mostly degraded. At 6 weeks, the constructs were much thinner in experimental group than those in control group 1 （ 1.44 ± 0.13mm vs 2.55 ± 0. 18mm in diameter ）. TEM showed periodical strata of collagen fibers in the constructs from experimental group and control group 1. However, histology in experimental group revealed longitudinal alignment of collagen fibers, which more resembled natural tendon than neotendon formed in control group 1. Besides, the maximum load to failure（ Newton/mm^2 ） was greater in experimental group than that in control group 1 （1. 107 ±0. 327 vs 0. 294 ± 0. 138, P 〈0.05）. Conclusion It＇ s possible to engineer tendon substitutes in vitro. Cyclic strain generated by a bioreactor may be the optimal mechanical stimulation and is currently under investigation.

Studies of nanoparticle delivery with in vitro bio-engineered microtissues

A variety of engineered nanoparticles,including lipid nanoparticles,polymer nanoparticles,gold nanoparticles,and biomimetic nanoparticles,have been studied as delivery vehicles for biomedical applications.When assessing the efficacy of a nanoparticle-based delivery system,in vitro testing with a model delivery system is crucial because it allows for real-time,in situ quantitative transport analysis,which is often difficult with in vivo animal models.The advent of tissue engineering has offered methods to create experimental models that can closely mimic the 3D microenvironment in the human body.This review paper overviews the types of nanoparticle vehicles,their application areas,and the design strategies to improve delivery efficiency,followed by the uses of engineered microtissues and methods of analysis.In particular,this review highlights studies on multicellular spheroids and other 3D tissue engineering approaches for cancer drug development.The use of bio-engineered tissues can potentially provide low-cost,high-throughput,and quantitative experimental platforms for the development of nanoparticle-based delivery systems.

Inkjet 3D bioprinting for tissue engineering and pharmaceutics

3D bioprinting has the capability to create 3D cellular constructs with the desired shape using a layer-by-layer approach.Inkjet 3D bioprinting,as a key component of 3D bioprinting,relies on the deposition of cell-laden droplets to create native-like tissues/organs which are envisioned to be transplantable into human body for replacing damaged ones.Benefiting from its superiorities such as high printing resolution and deposition accuracy,inkjet 3D bioprinting has been widely applied to various areas,including,but not limited to,tissue engineering and drug screening in pharmaceutics.Even though inkjet 3D bioprinting has proved its feasibility and versatility in various fields,the current applications of inkjet 3D bioprinting are still limited by the printing technique and material selection.This review,which specifically focuses on inkjet 3D bioprinting,firstly summarizes the techniques,materials,and applications of inkjet 3D bioprinting in tissue engineering and drug screening,subsequently discusses the major challenges that inkjet 3D bioprinting is facing,and lastly summarizes potential solutions to those challenges.