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The effect of liquid nitrogen exposure on the proliferative phase of Achilles tendon healing in Rattus norvegicus rats

By
Istan Irmansyah Irsan Orcid logo ,
Istan Irmansyah Irsan
Contact Istan Irmansyah Irsan

Department of Orthopaedic and Traumatology, Faculty of Medicine, Universitas Brawijaya, RSUD Prof. Dr. Saiful Anwar Malang , Malang , Indonesia

Edi Mustamsir Orcid logo ,
Edi Mustamsir

Department of Orthopaedic and Traumatology, Faculty of Medicine, Universitas Brawijaya, RSUD Prof. Dr. Saiful Anwar Malang , Malang , Indonesia

Panji Sananta Orcid logo ,
Panji Sananta

Department of Orthopaedic and Traumatology, Faculty of Medicine, Universitas Brawijaya, RSUD Prof. Dr. Saiful Anwar Malang , Malang , Indonesia

Satria Pandu Persada Isma Orcid logo ,
Satria Pandu Persada Isma

Department of Orthopaedic and Traumatology, Faculty of Medicine, Universitas Brawijaya, RSUD Prof. Dr. Saiful Anwar Malang , Malang , Indonesia

Dandy Drestanto Adiwignyo ,
Dandy Drestanto Adiwignyo

Department of Orthopaedic and Traumatology, Faculty of Medicine, Universitas Brawijaya, RSUD Prof. Dr. Saiful Anwar Malang , Malang , Indonesia

Hudzaifah Al Azmi Manaf ,
Hudzaifah Al Azmi Manaf

Department of Orthopaedic and Traumatology, Faculty of Medicine, Universitas Brawijaya, RSUD Prof. Dr. Saiful Anwar Malang , Malang , Indonesia

Adytia Andreyta Refa
Adytia Andreyta Refa

Department of Orthopaedic and Traumatology, Faculty of Medicine, Universitas Brawijaya, RSUD Prof. Dr. Saiful Anwar Malang , Malang , Indonesia

Editor: SELMA UZUNOVIĆ

Abstract

Aim Tendon healing involves a crucial proliferative phase, during which fibroblasts and fibrocytes orchestrate collagen deposition. The use of liquid nitrogen (LN) in orthopedic oncology may inadvertently affect adjacent tendon tissues. This study aimed to evaluate the impact of LN exposure on the histological features of tendon healing.

Methods This experimental study employed a randomized post-test-only control group design involving 24 males Rattus norvegicus, randomly divided into four groups: control (no LN exposure) and three treatment groups exposed to LN for 1, 5, and 10 minutes, respectively, following Achilles tendon transection and repair. After a 21-day healing period, histological analysis was performed to assess the counts of fibroblasts, fibrocytes, and collagen content. Statistical analyses included one-way ANOVA, Post-hoc Tukey, and Pearson correlation (p<0.05 was considered significant).

Results LN exposure significantly reduced fibroblast, fibrocyte, and collagen levels compared to controls (p<0.05). The 10-minute group showed the lowest counts. A significant negative correlation was found between LN immersion duration and the number of fibroblasts (r= -0.87), fibrocytes (r= -0.829), and collagen content (r= -0.83) (p<0.05).

Conclusion Liquid nitrogen (LN) impairs tendon healing in a dose-dependent manner, likely due to cryo-induced cell death and disruption of blood flow. This results in an acellular and avascular tendon matrix, hindering the repair process. LN exposure negatively impacts the proliferative phase of tendon healing in rats, suggesting the need for caution in clinical use to prevent damage to surrounding tendinous tissues.

Author Contributions

Conceptualization, I.I.I., H.A.A.M. and A.A.R.; Methodology, I.I.I., E.M., P.S., S.P.P.I. and D.D.A.; Supervision, I.I.I., E.M., P.S., S.P.P.I. and D.D.A.; Validation, I.I.I., E.M., P.S., S.P.P.I. and D.D.A.; Writing – original draft, I.I.I., H.A.A.M. and A.A.R.; Writing – review & editing, E.M., P.S., S.P.P.I. and D.D.A.; Data curation, H.A.A.M. and A.A.R.; Formal Analysis, H.A.A.M. and A.A.R. All authors have read and agreed to the published version of the manuscript.

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Funding Statement

No specific funding was received for this study.

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