Analysis and 3D Imaging of Morphometric - Mineral Structure Changes in Skulls of Male and Female Guinea Pigs During and After Developmental Period

SUMMARY: Even though morphometric and mineral studies related to the guinea pig (Cavia porcellus) skull have been carried out, this study is the first attempt to evaluate all developmental stages of male and female guinea pigs. This study aims to this study is to create 3D modeling of CT images obtained from the skulls of male and female guinea pigs during the developmental period (prepuberty and the period between puberty and adulthood) and following periods (young adulthood and old adulthood), to analyze some biometric bone data such as volume, surface area and length, and to assess the developmental analysis of the mineral matter change in their skulls. The CT-scanned skulls were transferred to 3D Slicer (5.0.2), which is used for 3D modeling. The surface area and volume were calculated by measuring the measurement points on the models. In addition, the XRF device was used to show elemental ratio changes during different developmental stages. According to metric measurements, a gradual increase was observed during the life period. The metric measurements of the skull bone had a higher measurement value in male guinea pigs than in their female counterparts. While Ca/P ratio increased up to the third group and partially decreased in the fourth group in males, it gradually increased from the first group to the fourth group in females. This study revealed that puberty, adulthood and sex were effective in the physical and chemical characterization of skull bone structure in guinea pigs.

Aunque se han realizado estudios morfométricos y de minerales relacionados con el cráneo del cobayo (Cavia porcellus), esta investigación es el primer intento de evaluar las etapas de desarrollo de cobayos machos y hembras. El objetivo de este estudio fue crear un modelado 3D de imágenes de tomografía computarizada obtenidas de los cráneos de cobayos machos y hembras durante el período de desarrollo (prepubertad y el período entre la pubertad y la edad adulta) y los períodos siguientes (edad adulta joven y edad adulta mayor), para analizar algunos datos biométricos de los huesos, como el volumen, la superficie y la longitud, y además, analizar el cambio de materia mineral en sus cráneos durante el desarrollo. Los cráneos escaneados se transfirieron a 3D Slicer (5.0.2), que se utiliza para el modelado 3D. El área de superficie y el volumen se calcularon midiendo los puntos de medición en los modelos. Además, se utilizó el dispositivo XRF para mostrar los cambios en las proporciones elementales durante diferentes etapas de desarrollo. Según mediciones métricas, se observó un aumento gradual durante el período de vida. Las medidas métricas del hueso del cráneo tuvieron un valor de medición más alto en los cobayos machos que en las hembras. Mientras que la relación Ca/P aumentó hasta el tercer grupo y disminuyó parcialmente en el cuarto grupo en los machos y aumentó gradualmente del primer grupo al cuarto grupo en las hembras. Este estudio reveló que la pubertad, la edad adulta y el sexo fueron efectivos en la caracterización física y química de la estructura ósea del cráneo en cobayos.

Physico-chemical and Histomorphometric Evaluation of Zinc-containing Hydroxyapatite in Rabbits Calvaria

Abstract The aim of this study was to characterize the physico-chemical properties and bone repair after implantation of zinc-containing nanostructured porous hydroxyapatite scaffold (nZnHA) in rabbits' calvaria. nZnHA powder containing 2% wt/wt zinc and stoichiometric nanostructured porous hydroxyapatite (nHA - control group) were shaped into disc (8 mm) and calcined at 550 °C. Two surgical defects were created in the calvaria of six rabbits (nZnHA and nHA). After 12 weeks, the animals were euthanized and the grafted area was removed, fixed in 10% formalin with 0.1 M phosphate buffered saline and embedded in paraffin (n=10) for histomorphometric evaluation. In addition, one sample from each group (n=2) was embedded in methylmethacrylate for the SEM and EDS analyses. The thermal treatment transformed the nZnHA disc into a biphasic implant composed of Zn-containing HA and Zn-containing β-tricalcium phosphate (ZnHA/βZnTCP). The XRD patterns for the nHA disc were highly crystalline compared to the ZnHA disc. Histological analysis revealed that both materials were biologically compatible and promoted osteoconduction. X-ray fluorescence and MEV-EDS of nZnHA confirmed zinc in the samples. Histomorphometric evaluation revealed the presence of new bone formation in both frameworks but without statistically significant differences (p>0.05), based on the Wilcoxon test. The current study confirmed that both biomaterials improve bone repair, are biocompatible and osteoconductive, and that zinc (2wt%) did not increase the bone repair. Additional in vivo studies are required to investigate the effect of doping hydroxyapatite with a higher Zn concentration.

Resumo O objetivo deste estudo foi caracterizar físico-químicamente e avaliar o reparo ósseo de discos de hidroxiapatita porosa contendo zinco após a implantação em calvária de coelhos (nZnHA). O pó de nZnHA e o pó hidroxiapatita porosa estequiométrica nanoestruturada (controle - nHA) foram confeccionados em discos (8 mm) e calcinados a 550˚C. Dois defeitos cirúrgicos foram criados na calvária de seis coelhos para a implantação dos discos. Após 12 semanas, os animais foram eutanasiados e as áreas enxertadas foram removidas, fixadas em formol a 10% e embebidas em parafina (n=10) para avaliação histomorfométrica. Além disso, uma amostra de cada grupo (n=2) foi embebida em metilmetacrilato para análise de MEV e EDS. O tratamento térmico dos discos de nZnHA transformou-os em implantes bifásicos compostos por HA contendo Zinco e β fosfato tricálcico com Zinco (ZnHA/βZnTCP). Os discos de nHA, apresentaram-se altamente cristalinos e com baixa solubilidade quando comparados aos discos de ZnHA. A análise histológica revelou que ambos os materiais foram biologicamente compatíveis e promoveram a osteocondução. As análises de FRX e MEV-EDS confirmaram a presença do zinco nas amostras de nZnHA. A avaliação histomorfométrica revelou a presença de neoformação óssea em ambos os grupos, porém sem diferenças estatísticas entre eles, com base no teste de Wilcoxon (p>0,05). O presente estudo confirmou que ambos os biomateriais otimizaram o reparo ósseo, foram biocompatíveis e osseocondutivos e a presença do zinco não favoreceu o reparo ósseo. Estudos adicionais in vivo devem ser conduzidos a fim de investigar o efeito de maiores concentrações de zinco.