Sains Malaysiana 53(12)(2024): 3349-3364

http://doi.org/10.17576/jsm-2024-5312-18

Cytoprotective Role of Chondrocyte Coculturing for Enhanced Cartilage Regeneration

(Peranan Sitopelindung Pengkulturan Kondrosit untuk Penjanaan Semula Rawan Dipertingkat)

MUHAMMAD FAIZAN TARIQ¹, SUMERA RASHID¹, AISHA TARAR¹, UMAR SAJJAD¹, MUHAMMAD RAUF AHMED², BUSHRA IJAZ¹, HAIBA KAUL³ & NOREEN LATIEF^1,*

¹National Centre of Excellence in Molecular Biology, University of The Punjab, Lahore, Pakistan
²Department of Molecular Biology and Biochemistry, Shaheed Zulfiqar Ali Bhutto Medical University, Islamabad, Pakistan
³Department of Animal Breeding and Genetics, University of Veterinary and Animal Sciences, Lahore, Pakistan

Diserahkan: 28 Julai 2024/Diterima: 27 September 2024

Abstract

Osteoarthritis is characterized by the progressive deterioration of articular cartilage, leading to joint pain and functional impairments. Current treatment options are limited in their ability to stimulate cartilage regeneration. Evidence suggests that the co-culture technique, involving the interplay of multiple cell types, may effectively restore damaged cartilage. This investigation assessed the regenerative impact of co-culture on osteoarthritis-affected rat knee joints. Gene expression profiling validated phenotypic and biochemical expression analysis. Compared to the control group, the co-cultivated cohorts showed elevated levels of cartilage-specific markers, such as collagen and aggrecan. Notably, the group with stress and co-cultured with normal and osteoarthritic chondrocytes (H₂O₂+N+OA) demonstrated significant results, including lower LDH release (15.06 ± 1.461), decreased glycosaminoglycan levels (1.551 ± 0.1487), and reduced cell death percentage (17.50 ± 3.536) compared to the H₂O₂ control. Safranin-O staining retention also increased (28.89 ± 2.846), indicating enhanced cartilage matrix retention. Enhanced expression of survival markers such as Bcl (0.3974 ± 0.02241) and decreased apoptotic markers like Bax (0.2961 ± 0.01199) were observed, confirming the stimulation of critical genes involved in cartilage development and matrix synthesis. These findings support the potential of co-culture technology to accelerate cartilage regeneration and offer an innovative strategy to impede osteoarthritis progression.

Keywords: Cartilage regeneration; cellular survival; co-culturing; osteoarthritis

Abstrak

Osteoartitis dicirikan oleh kemerosotan progresif rawan artikular yang membawa kepada sakit sendi dan gangguan fungsi. Pilihan rawatan semasa adalah terhad dalam keupayaannya untuk merangsang pertumbuhan semula rawan. Bukti menunjukkan bahawa teknik pengkulturan bersama yang melibatkan interaksi pelbagai jenis sel boleh memulihkan rawan yang rosak dengan berkesan. Penyelidikan ini menilai kesan penjanaan semula pengkulturan bersama pada sendi lutut tikus yang terjejas oleh osteoartitis. Pemprofilan pengekspresan gen mengesahkan analisis pengekspresan fenotip dan biokimia. Berbanding dengan kumpulan kawalan, kohort yang diusahakan bersama menunjukkan peningkatan tahap penanda khusus rawan, seperti kolagen dan aggrecan. Terutamanya, kumpulan dengan tekanan dan pengkulturan bersama dengan kondrosit normal dan kondrosit osteoartitis (H₂O₂+N+OA) menunjukkan hasil yang ketara, termasuk pelepasan LDH yang lebih rendah (15.06 ± 1.461), penurunan paras glikosaminoglikan (1.551 ± 0.1487) dan pengurangan peratusan kematian sel (17.50 ± 3.536) berbanding kawalan H₂O₂. Pengekalan pewarnaan Safranin-O juga meningkat (28.89 ± 2.846), menunjukkan pengekalan matriks rawan dipertingkat. Pengekspresan penanda kelangsungan hidup yang dipertingkatkan seperti Bcl (0.3974 ± 0.02241) dan penurunan penanda apoptosis seperti Bax (0.2961 ± 0.01199) telah diperhatikan, mengesahkan rangsangan gen kritikal yang terlibat dalam pembangunan rawan dan sintesis matriks. Penemuan ini menyokong potensi teknologi pengkulturan bersama untuk mempercepatkan pertumbuhan semula rawan dan menawarkan strategi inovatif untuk menghalang perkembangan osteoartitis.

Kata kunci: Kelangsungan hidup sel; osteoartitis; pengkulturan bersama; penjanaan semula rawan

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*Pengarang untuk surat-menyurat; email: noreen.latief@cemb.edu.pk