Measurement of Three-Dimensional Anisotropic Diffusion by Multiphoton Fluorescence Recovery after Photobleaching

Shi, Changcheng; Cisewski, Sarah E.; Bell, P. Darwin; Yao, Hai

doi:10.1007/s10439-013-0939-7

Cited by 13 publications

(19 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We suggest that for initial EO to converge into a spatially well-controlled process, the gradual diffusion of signaling molecules towards BMSCs in the pre-cartilage region is required to induce and orchestrate their proliferation and chondrogenic differentiation 59 . This is supported by theoretical and experimental work that showed facilitated diffusion along aligned fiber networks compared with perpendicular or random structures 60 , 61 . Regarding the late events in the EO process, terminal chondrocyte hypertrophy and matrix mineralization are coupled to vascularization.…”

Section: Discussionmentioning

confidence: 53%

A biomaterial with a channel-like pore architecture induces endochondral healing of bone defects

et al. 2018

View full text Add to dashboard Cite

Biomaterials developed to treat bone defects have classically focused on bone healing via direct, intramembranous ossification. In contrast, most bones in our body develop from a cartilage template via a second pathway called endochondral ossification. The unsolved clinical challenge to regenerate large bone defects has brought endochondral ossification into discussion as an alternative approach for bone healing. However, a biomaterial strategy for the regeneration of large bone defects via endochondral ossification is missing. Here we report on a biomaterial with a channel-like pore architecture to control cell recruitment and tissue patterning in the early phase of healing. In consequence of extracellular matrix alignment, CD146+ progenitor cell accumulation and restrained vascularization, a highly organized endochondral ossification process is induced in rats. Our findings demonstrate that a pure biomaterial approach has the potential to recapitulate a developmental bone growth process for bone healing. This might motivate future strategies for biomaterial-based tissue regeneration.

show abstract

Section: Discussionmentioning

confidence: 53%

A biomaterial with a channel-like pore architecture induces endochondral healing of bone defects

et al. 2018

View full text Add to dashboard Cite

show abstract

“…As the speed and sensitivity of confocal microscopes is ever increasing, 3D image acquisition of the recovery process may become within reach as well, which will lead to a better view of the recovery process along the optical axis and hence a better interpretation of the data, as was recently demonstrated (Shi et al . 2014). Equally important is a continuous development of new and better FRAP probes that can be altered in molecular weight, shape, charge, polarity, hydrophobicity, etc.…”

Section: Conclusion and Future Outlookmentioning

confidence: 99%

Fluorescence recovery after photobleaching in material and life sciences: putting theory into practice

Lorén¹,

Hagman²,

Jonasson

et al. 2015

Quart. Rev. Biophys.

134

121

View full text Add to dashboard Cite

Fluorescence recovery after photobleaching (FRAP) is a versatile tool for determining diffusion and interaction/binding properties in biological and material sciences. An understanding of the mechanisms controlling the diffusion requires a deep understanding of structure-interaction-diffusion relationships. In cell biology, for instance, this applies to the movement of proteins and lipids in the plasma membrane, cytoplasm and nucleus. In industrial applications related to pharmaceutics, foods, textiles, hygiene products and cosmetics, the diffusion of solutes and solvent molecules contributes strongly to the properties and functionality of the final product. All these systems are heterogeneous, and accurate quantification of the mass transport processes at the local level is therefore essential to the understanding of the properties of soft (bio)materials. FRAP is a commonly used fluorescence microscopy-based technique to determine local molecular transport at the micrometer scale. A brief high-intensity laser pulse is locally applied to the sample, causing substantial photobleaching of the fluorescent molecules within the illuminated area. This causes a local concentration gradient of fluorescent molecules, leading to diffusional influx of intact fluorophores from the local surroundings into the bleached area. Quantitative information on the molecular transport can be extracted from the time evolution of the fluorescence recovery in the bleached area using a suitable model. A multitude of FRAP models has been developed over the years, each based on specific assumptions. This makes it challenging for the non-specialist to decide which model is best suited for a particular application. Furthermore, there are many subtleties in performing accurate FRAP experiments. For these reasons, this review aims to provide an extensive tutorial covering the essential theoretical and practical aspects so as to enable accurate quantitative FRAP experiments for molecular transport measurements in soft (bio)materials.

show abstract

“…Nonfixed, nondecalcified maxillae freshly removed from mice were rinsed and then soaked in 50% glycerol for 1 hour . Jaws were mounted in optimal cutting temperature (OCT) and sliced to obtain a flat‐exposed longitudinal slice of PDL using a sledge microtome (Leica Microsystems).…”

Section: Methodsmentioning

confidence: 99%

Inhibition of transglutaminase activity in periodontitis rescues periodontal ligament collagen content and architecture

Rosset

Trombetta-eSilva

Hepfer

et al. 2019

J of Periodontal Research

Self Cite

View full text Add to dashboard Cite

Background and objective Periodontal disease (PD) afflicts approximately 50% of the population in the United States and is characterized by chronic inflammation of the periodontium that can lead to loss of the periodontal ligament through collagen degradation, loss of alveolar bone, and to eventual tooth loss. Previous studies have implicated transglutaminase (TG) activity in promoting thin collagen I fiber morphology and decreased mechanical strength in homeostatic PDL. The aim of this study was to determine whether TG activity influenced collagen assembly in PDL in the setting of periodontal disease. Material and methods A ligature model was used to induce clinically relevant PD in mice. Mice with ligature were assessed at 5 and 14 days to determine PDL collagen morphology, transglutaminase (TG) activity, and bone loss. The effects of inhibition of TG on PDL were assessed by immunohistochemistry and second‐harmonic generation (SHG) to visualize collagen fibers in native tissue. Results Ligature placement around the 2nd molar resulted in significant bone loss and a decrease in total collagen content after 5 days of ligature placement. A significant increase in thin over thick fibers was also demonstrated in mice with ligature at 5 days associated with apparent increases in immunoreactivity for TG2 and for TG‐mediated N‐ε‐γ‐glutamyl cross‐links in PDL. Inhibition of TG activity increased total collagen and thick collagen fiber content over vehicle control in mice with ligature for 5 days. SHG of PDL was used to visualize and quantify the effects of TG inhibition on enhanced collagen fiber organization in unfixed control and diseased PDL. Conclusion These studies support a role of TG in regulating collagen fiber assembly and suggest that strategies to inhibit TG activity in disease might contribute to restoration of PDL tissue integrity.

show abstract

Measurement of Three-Dimensional Anisotropic Diffusion by Multiphoton Fluorescence Recovery after Photobleaching

Cited by 13 publications

References 30 publications

A biomaterial with a channel-like pore architecture induces endochondral healing of bone defects

A biomaterial with a channel-like pore architecture induces endochondral healing of bone defects

Fluorescence recovery after photobleaching in material and life sciences: putting theory into practice

Inhibition of transglutaminase activity in periodontitis rescues periodontal ligament collagen content and architecture

Contact Info

Product

Resources

About