Impacts of aging on murine cartilage biomechanics and chondrocyte in situ calcium signaling

Fan, Mingyue; Wang, Chao; Kwok, Bryan; Kahle, Elizabeth R.; He, Lu; Lü, Xin; Mauck, Robert L.; Han, Lin

doi:10.1016/j.jbiomech.2022.111336

Cited by 3 publications

(5 citation statements)

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“…Previous studies have shown the hypo-osmolar conditions increase the calcium signaling response in in-situ chondrocytes via the TRPV4 channel activation 91,178 Inducing GCaMP expression in mature in-situ axolotl chondrocytes is challenging without inducing significant tissue damage due to overheating. Hence, to study calcium signaling in chondrocytes under in-vivo conditions, a transgenic animal with a calcium reporter (GCaMP) linked to active SOX-9 expression will need to be developed.…”

Section: Discussionmentioning

confidence: 99%

“…We propose to use GCaMP in regenerating axolotl limb tissues to study the calcium signaling dynamics in-vivo. However, previous studies, including ours, explored calcium signaling in chondrocytes in bovine cartilage explants using chemical indicators 7,91,178 . In order to compare the differences between signaling characteristics observed in axolotl cartilage (an amphibian) with the existing literature on bovine cartilage (mammalian system), a systematic approach is required.…”

Section: Gap In Knowledge and Motivationmentioning

confidence: 95%

“…Additionally, cells in explants respond immediately upon loading 80,81 while cells embedded in 3D constructs respond with a delay of hundreds of seconds 89 . Similarly, various physical stimuli such as deformation, osmolarity, and strain elicit increased calcium influx in chondrocytes, with greater stimulus magnitudes resulting in a higher percentage of cells signaling 7,8,90,91 . These studies underscore the significance of both the matrix environment and load characteristics in influencing intracellular calcium signaling in chondrocytes.…”

Section: Extracellular Matrix Affects Cellular Responsementioning

confidence: 99%

“…As discussed in chapter 3, calcium signaling in chondrocytes is highly dependent on its matrix environment 7,81,85,121 . While numerous studies have explored calcium dynamics in in-situ chondrocytes under in-vitro conditions 7,80,91,129,178 or in developing limb bud micro mass cell cultures 177 , examining chondrocytes in an in-vivo setting remains challenging due to the inherent difficulties in accessing tissues for in-vivo imaging. To address this, an ideal animal model is needed-one that is small enough to fit under a microscope for observation and one that offers a large, easily accessible surface area.…”

Section: Effect Of Matrix Environment and Choice Of Animal Modelmentioning

confidence: 99%

“…Although murine models have previously been used in cartilage studies 178 , challenges arise due to the presence of secondary ossification centers 180 , making the articular cartilage difficult to image in-vivo under a microscope 181 . Axolotls, with their regenerative features and accessible chondro-epiphyses (see Figure 5.1), present a unique opportunity to study calcium signaling dynamics not only in mature articular cartilage but also in regenerating limb tissues.…”

Section: Effect Of Matrix Environment and Choice Of Animal Modelmentioning

confidence: 99%

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Cartilage mechanobiology during limb growth

Kondiboyina

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The mechanobiology of cartilage during limb growth represents a complex interplay between mechanical forces and biological processes. However, the fundamental processes that involve the formation of cartilage from mesenchymal stem cells are not fully understood. Further, the cellular level response of cartilage in its native environment under physiological load is not fully characterized. This thesis aims to bridge critical gaps in our understanding of limb development and regeneration by investigating the nuanced interactions between mechanics and cellular processes within cartilaginous tissues.Identifying these research gaps, this thesis encompasses three specific aims. Aim 1 focuses on characterizing the viscoelastic material properties of growing limbs, employing axolotls as an animal model. Our results reveal significant increases in both instantaneous and equilibrium shear moduli during limb regeneration, coupled with notable changes in short-and long-term stress relaxation times. The glycosaminoglycan content also increases during development. Aim 2 explores the calcium signaling response of in-situ chondrocytes under physiologically relevant cyclic loads and dynamic hydrostatic pressure. Our findings underscore a strain rate-dependent increase in the percentage of responsive cells under compressive loads, with non-distinct time characteristics across loading conditions. Conversely, low magnitude dynamic hydrostatic pressure showed no significant impact on calcium signaling in chondrocytes. Aim 3 investigates the expression of mechanosensitive ion channels (TRPV4, PIEZO1, and PIEZO2) during axolotl limb regeneration. The study unveils the presence of TRPV4 and PIEZO2 in blastemal cells during early and late regeneration, with heightened expression in the condensing mesenchyme during late regeneration.These findings taken together shed light on the dynamic changes in the mechanical environment during limb growth and the intricate interplay between mechanics and cellular responses. The implications of abnormal mechanobiological processes are profound, contributing to developmental disorders and musculoskeletal diseases. Understanding these fundamental mechanisms under physiological conditions opens avenues for therapeutic strategies aimed at promoting proper limb development and mitigating skeletal abnormalities. Future research will focus on elucidating the functional roles of mechanosensitive channels during regeneration and further expanding our understanding of the complex interconnections between mechanics and skeletal biology. iii I am deeply grateful to my colleagues at the Shefelbine lab, past, and present, whose support and camaraderie have been invaluable. Quentin Meslier, Soha Ben Tahar and Lindsey Young, thank you. Special acknowledgment goes to Noah Mooney for his remarkable work on engineering the hydrostatic pressure system. Working alongside him was truly enjoyable. iv Special thanks to Dr. Helen Markewich for training me in essential laboratory skills. I extend my appreciation to the support...

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Section: Discussionmentioning

confidence: 99%

Section: Gap In Knowledge and Motivationmentioning

confidence: 95%