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Utilizing tissue clearing based 3-D imaging to quantitatively study neural regulation of craniofacial mesenchymal stem cells

Award Number: R21DE027928
ORGANIZATION: NATIONAL INSTITUTE OF DENTAL AND CRANIOFACIAL RESEARCH
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

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Utilizing tissue clearing based 3-D imaging to quantitatively study neural regulation of craniofacial mesenchymal stem cells Recent discoveries of bone regulation by the nervous system has led to the identification of new regulatory axes between the nervous system and bone1-3. Various studies indicate that signal from the central or peripheral nervous system directly or indirectly regulates the bone remodeling process1, 2. One technical challenge on studying the nerve-bone interactions is to visualize nerve fibers on or within bones because of their fine size (~1µm diameter) and the distant location of the neuron cell bodies. In addition, 2-D information obtained from sections made it difficult or impossible to quantify spatial interactions between complex craniofacial tissues. Development of 3-D imaging based tissue clearing technique provides solutions to these two issues. Tissue clearing technique is a recent breakthrough in the neuroscience field. By rendering tissue transparent, 3-D images of the whole organ can be achieved directly with a confocal microscope without sectioning4. We recently developed a new tissue clearing technique named PEG Associated Solvent System (PEGASOS). The PEGASOS method efficiently clears both hard and soft tissue to high transparency and protects endogenous fluorescence with no loss. The PEGASOS method was initially developed to study the neuron connectome within the brain (manuscript in revision). Pilot experiments suggested that it can also be applied on the craniofacial hard tissue. Skull bone, teeth and femur harvested from adult mice can be cleared to nearly invisible after treatment without losing GFP fluorescence. We were able to acquire the 3-D images of vascular network within the tooth, suture, dura and calvarial bone. We were also able to acquire the 3-D images of nerves within the femur periosteum, tibia bone marrow, suture and periodontal ligamental space. Quantitative analysis of these 3-D images indicated that vasculatures and nerves within craniofacial hard tissue present highly heterogenous distribution pattern. In contrast to previous study, our preliminary results suggested that Gli1+ cells within the suture are closely associated with the vasculatures. In the current proposal, we propose to improve the PEGASOS method for craniofacial hard tissue research. With the spatial information provided by PEGASOS based 3-D imaging, we would like to quantitatively test the hypothesis that craniofacial MSCs are associated with the neurovascular bundle (NVB) and nerves are essential for stem cell migration towards the injury site


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2023 ( Subtotal = -$49 )
2023	2019	TEXAS A & M UNIVERSITY SYSTEM HEALTH SCIENCE CENTER	6003 TAMU	COLLEGE STATION	TX	77843	BRAZOS	USA	Oral Diseases and Disorders Research	000	2	11/21/2022	NON-COMPETING CONTINUATION	-$49
														Subtotal = -$49

Issue Date FY: 2022 ( Subtotal = -$18,791 )
2022	2019	TEXAS A&M UNIVERSITY SYSTEM, THE	8441 RIVERSIDE PARKWAY BLDG 1 STE 3100	BRYAN	TX	77807	BRAZOS	USA	Oral Diseases and Disorders Research	000	2	10/13/2021	NON-COMPETING CONTINUATION	$0
2022	2019	TEXAS A&M UNIVERSITY SYSTEM, THE	8441 RIVERSIDE PARKWAY BLDG 1 STE 3100	BRYAN	TX	77807	BRAZOS	USA	Oral Diseases and Disorders Research	001	2	1/19/2022	NON-COMPETING CONTINUATION	-$18,791
														Subtotal = -$18,791

Issue Date FY: 2019 ( Subtotal = $185,625 )
2019	2019	TEXAS A&M UNIVERSITY SYSTEM, THE	8441 RIVERSIDE PARKWAY BLDG 1 STE 3100	BRYAN	TX	77807	BRAZOS	USA	Oral Diseases and Disorders Research	000	2	7/26/2019	NON-COMPETING CONTINUATION	$185,625
														Subtotal = $185,625

Issue Date FY: 2018 ( Subtotal = $222,750 )
2018	2018	TEXAS A&M UNIVERSITY SYSTEM,THE	200 TECHNOLOGY WAY, STE 2079	COLLEGE STATION	TX	77845	BRAZOS	USA	Oral Diseases and Disorders Research	000	1	8/10/2018	NEW	$222,750
														Subtotal = $222,750

Grand Total All Awards = $389,534

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Utilizing tissue clearing based 3-D imaging to quantitatively study neural regulation of craniofacial mesenchymal stem cells

Award Number: R21DE027928

ORGANIZATION: NATIONAL INSTITUTE OF DENTAL AND CRANIOFACIAL RESEARCH

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

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