EFEK GEN CARBONIC ANHYDRASE 12 (CA12) PADA STRUKTUR MAKROSKOPIS, MIKROARSITEKTUR, DAN HISTOLOGIS MENCIT JANTAN DENGAN DELESI SPESIFIK GEN CA12

Abstract

Introduction: Carbonic Anhydrase 12 (CA12) gene is responsible for the production of Carbonic Anhydrase XII protein (CAXII). CAXII is a major catalyst for converting carbon dioxide (CO2) and water (H2O) into bicarbonate ion (HCO3-) and hydrogen (H+) which responsible for various physiological properties. This study aims to analyze the effect of CA12 deletion on bone growth, microarchitecture and histological structure of the calvarial.

Methods: Study was conducted on 16 weeks old male mice divided into 2 groups; Osteocyte Carbonic Anhydrase 12 floxed mice (ObCar12 HOM-C) and osteocyte specific knock-out of CA12 gene (ObCar12 HOM-KO). Macroscopic structures of the calvarias were analyzed using faxitron images and measured using Image J. Calvarial microarchitecture were analyzed using micro-CT Skyscan 1174 and CT-Analyses while histological assessments were conducted on Hematoxylin & Eosin (H&E) and Renee’s modification of Toluidine Blue (TOL) staining and imaged using Nanozoomer and NDP analyzer. Results were analyzed using t-test, and p is set at 0.05.

Results: There is no significant effect of specific gene deletion on the growth of calvarias shown by similar calvarial width, length, and depth (0.68 ± 0.01 vs 0.68 ± 0.01) between two groups. Calvarial microarchitecture of ObCar12 also shows no significant differences in BV/TV (5.33 ± 0.47 vs 5.01 ± 0.57), trabecular number, thickness or separation. Histological analyses of the calvarias by measuring occupied lacunae (90.0 ± 0.83 vs 91.7 ± 1.42), empty lacunae, average number of lacunae, and total area of lacunae in both groups also shows no significant differences (p>0.05).

Conclusion: Specific deletion of CA12 gene in osteocytes does not affect calvarias macroscopic, microarchitecture and histological structures.