Interactions among osteoblastic cells, Staphylococcus aureus, and chitosan-immobilized titanium implants in a postoperative coculture system: An in vitro study.

Biomaterial-related infections (BRIs) have become a major challenge in the field of orthopedic implants. In this study, we delved into the problem of BRI and attempted to reduce the possibility of BRI incidence via surface modification of titanium (Ti) with chitosan (SA-CS-Ti). To comprehensively evaluate the anti-infection potential of SA-CS-Ti, we first constructed a postoperative infection (POI) model with varying concentrations of bacteria (102  CFU/sample and 104  CFU/sample) and a constant number of SaOS-2 cells (105 /sample). Then, we biologically characterized the interactions between the SaOS-2 cells, bacteria, and different Ti implants using the POI model. The results from the osteoblastic cell and bacterial attachment tests demonstrated that the SA-CS-Ti surfaces exhibit superior osteogenic behavior relative to other Ti surfaces studied while showing significant anti-infective activities in the POI model with a low infection ratio (bacteria: cell ratio of 0.001:1) 30 min after infection. Additionally, the SA-CS-Ti surfaces showed significantly reduced (p < 0.05) bacteria proliferation compared to the control Ti surfaces (UN-Ti), demonstrating their antifouling property. The significantly increased (p < 0.05) sensitivity of Staphylococcus. aureus adhered to the SA-CS-Ti surfaces against cefazolin (1 mg/L treatment) and gentamicin (10 mg/L and 100 mg/L treatment) in the coculture system augmented potential of SA-CS-Ti to be used as orthopedic implants. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 586-594, 2016.

Bacteria and osteoblast adhesion to chitosan immobilized titanium surface: A race for the surface.

In order to evaluate the anti-infective efficacy of the titanium implant materials, two co-culture systems, a low-bacteria/osteoblast (L-B) and a high-bacteria/osteoblast system (H-B), were established. Untreated (UN-Ti), sulfuric acid-treated (SA-Ti), and chitosan immobilized titanium (SA-CS-Ti) materials were developed and evaluated. Bacteria and osteoblast behaviors, including initial attachment (evaluated at 30 mins), adhesion (evaluated at 4 h), and osteoblast spreading on each material surface were evaluated using quantification assays, scanning electron microscopy (SEM), and confocal microscopy. Quantification analysis at 30 mins showed significantly higher number of osteoblast present on SA-CS-Ti in both L-B (10,083 ± 2626) and H-B (23,592 ± 2233) than those on the UN-Ti (p<0.05). SEM observation and confocal microscopy results showed more surface area was occupied by adhered osteoblasts on SA-CS-Ti than UN-Ti and SA-Ti in both co-culture systems at 30 mins. At all time points, SA-CS-Ti had the lowest level of bacterial adhesion compared to UN-Ti and SA-Ti in both co-culture systems. A significantly (p<0.05) lower number of bacteria were recovered from SA-CS-Ti (2233 ± 681) in the H-B system compared to UN-Ti (5367 ± 1662) and SA-Ti (4533 ± 680) at 4h. Quantitative and qualitative co-culture results show the great potential of chitosan immobilization onto implant materials to prevent implant-associated infections.

Chondroprotective supplementation promotes the mechanical properties of injectable scaffold for human nucleus pulposus tissue engineering.

A result of intervertebral disc (IVD) degeneration, the nucleus pulposus (NP) is no longer able to withstand applied load leading to pain and disability. The objective of this study is to fabricate a tissue-engineered injectable scaffold with chondroprotective supplementation in vitro to improve the mechanical properties of a degenerative NP. Tissue-engineered scaffolds were fabricated using different concentrations of alginate and calcium chloride and mechanically evaluated. Fabrication conditions were based on structural and mechanical resemblance to the native NP. Chondroprotective supplementation, glucosamine (GCSN) and chondroitin sulfate (CS), were added to scaffolds at concentrations of 0:0µg/mL (0:0-S), 125:100µg/mL (125:100-S), 250:200µg/mL (250:200-S), and 500:400µg/mL (500:400-S), GCSN and CS, respectively. Scaffolds were used to fabricate tissue-engineered constructs through encapsulation of human nucleus pulposus cells (HNPCs). The tissue-engineered constructs were collected at days 1, 14, and 28 for biochemical and biomechanical evaluations. Confocal microscopy showed HNPC viability and rounded morphology over the 28 day period. MTT analysis resulted in significant increases in cell proliferation for each group. Collagen type II ELISA quantification and compressive aggregate moduli (HA) showed increasing trends for both 250:200-S and the 500:400-S groups on Day 28 with significantly greater HA compared to 0:0-S group. Glycosaminoglycan and water content decreased for all groups. Results indicate the increased mechanical properties of the 250:200-S and the 500:400-S was due to production of a functional matrix. This study demonstrated potential for a chondroprotective supplemented injectable scaffold to restore biomechanical function of a degenerative disc through the production of a mechanically functional matrix.