Doxycycline preserves chondrocyte viability and function in human and calf articular cartilage ex vivo

Abstract

Prolonging chondrocyte survival is essential to ensure fresh osteochondral (OC) grafts for treatment of articular cartilage lesions. Doxycycline has been shown to enhance cartilage growth, disrupt terminal differentiation of chondrocytes, and inhibit cartilage matrix degradation. It is unknown whether doxycycline prolongs chondrocyte survival in OC grafts. We hypothesized that doxycycline protects against chondrocyte death and maintains function of articular cartilage. To test this hypothesis, we employed human and calf articular cartilages, and incubated chondrocytes isolated from cartilage or cartilage plugs with doxycycline (0, 1 or 10 μg/ml) at either 37°C or 4°C. Chondrocyte viability, apoptosis, glycosaminoglycan (GAG), collagen, and mechanical test in cartilage plugs were measured. We found that reduced chondrocyte viability, increased chondrocyte apoptosis, reduced GAG contents, and impaired equilibrium modulus in cartilage plugs were observed in a time‐dependent manner at both 37°C and 4°C. Chondrocyte viability was further reduced when the plugs were cultured at 4°C as compared to 37°C. Doxycycline prolonged viability and reduced apoptosis of chondrocytes during culture of cartilage plugs. Functionally, doxycycline protected against reduced production of GAG and collagen II as well as impaired mechanical properties in cartilage plugs during culture. Mechanistically, doxycycline increased mitochondrial respiration in cultured chondrocytes. In conclusion, preservation at 37°C is beneficial for maintaining chondrocyte viability in cartilage plugs compared to 4°C. Incubation of doxycycline protects against chondrocyte apoptosis, reduced extracellular matrix, and impaired mechanical properties in cartilage plugs. The findings provide a potential approach using doxycycline at 37°C to preserve chondrocyte viability in fresh OC grafts for treatment of articular cartilage lesions.

Review of Intra-Articular Use of Antibiotics and Antiseptic Irrigation and Their Systematic Association with Chondrolysis

“…Other studies did not demonstrate this same chondrotoxicity. 18,19 For instance, Yue et al 23 found that doxycycline decreased chondrocyte apoptosis, increased type II collagen synthesis, and increased mechanical integrity. Doxycycline has also been shown to protect the articular environment through decreased expression of catabolic cytokines.…”

Regeneration of partially decellularized tracheal scaffolds in a mouse model of orthotopic tracheal replacement

“…Chondrocyte viability plays a critical role in maintaining the biochemical and biomechanical properties necessary to prevent stenosis or collapse in tracheal replacement, however the preservation of chondrocyte viability remains a challenge in tracheal tissue engineering. 19,41,42 Modulation of decellularization protocol, such as optimization of decellularization time, can preserve viability of chondrocytes in DTS. 19 We also observed a higher mortality rate of hosts receiving DTS than that of STG control (p = 0.0490).…”

A Multimodal Approach to Quantify Chondrocyte Viability for Airway Tissue Engineering

“…Chondrocytes are responsible for the maintenance of the graft matrisome; the viability of these cells could prove to play a critical role in graft performance. [28][29][30] Beyond decellularization, graft viability is related to ischemia time and storage conditions; these factors can limit the creation of a donor tissue bank. Wellestablished approaches of cryopreservation methods to biobank organs have been developed to enhance viability.…”

“…Graft chondrocyte viability is now a new and important consideration. Chondrocytes are responsible for the maintenance of the graft matrisome; the viability of these cells could prove to play a critical role in graft performance 28–30 . Beyond decellularization, graft viability is related to ischemia time and storage conditions; these factors can limit the creation of a donor tissue bank.…”