Red propolis extract associated to platelet-rich plasma and stromal cells with focus in cell therapy and functional tissue regeneration

GARCIA, CHARLENE S.C.; GARCIA, PAULO MIGUEL C.; SANTOS, OTÁVIO B.A.F.; STEFFENS, DANIELA; MARTINS, SANDRO T.; PRANKE, PATRICIA; CRESPO, JANAÍNA S.; HENRIQUES, JOÃO ANTONIO P.; ROESCH-ELY, MARIANA

doi:10.1590/0001-3765202420240100

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HEALTH SCIENCES • An. Acad. Bras. Ciênc. 96 (suppl 1) • 2024 • https://doi.org/10.1590/0001-3765202420240100 linkcopy

Red propolis extract associated to platelet-rich plasma and stromal cells with focus in cell therapy and functional tissue regeneration

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Abstract

The use of platelet-rich plasma (PRP) and adipose-derived stromal cells (ADSC) have been investigated as a form of wound healing enhancement. The objective of this work was to evaluate the association of red propolis (RP) and PRP as inducers of ADSC for application in tissue regeneration. Adipose tissue post-collection and post-cryopreservation was isolated with type II collagenase, characterized by flow cytometry, and differentiated into osteogenic, chondrogenic and adipose cell. The viability of ADSC was evaluated when exposed to different concentrations of RP using the MTT and trypan blue assay. Acridine orange and ethidium bromide (AO/EB) was performed to evaluate cell death events. Horizontal migration methods were investigated in ADSC using autologous and homologous PRP associated with RP (PRP/RP). All assays were processed in triplicate. Flow cytometry and cellular differentiation showed that type II collagenase was effective for isolating ADSC post-collection and post-cryopreservation. RP extracts at concentrations of up to 50 μg.mL^-1 presented no cytotoxic effects. Association of PRP and RP at 25 and 50 μg.ml^-1 influenced ADSC migration, with total closure on the seventh day after exposition. The results here presented could stimulate proliferation of ADSC cells that may contribute directly or indirectly to the reconstructive process of tissue regeneration.

Key words
adipose derived stromal cells; cell therapy; platelet-rich plasma; red propolis; tissue regeneration

IC₅₀
Time	1 day	2 days	3 days	7 days
Concentration of RP µg.mL^-1	119.93±2.45	95.22±1.56	90.75±2.77	73.56±1.32

		25 µg	50 µg	100 µg	200 µg
		1 day
Cell count (%)	Viable	96.03±0.90	93.99±1.28	86.42±1.53	7.00±1.89
	Apoptosis	3.76±0.84	6.01±1.28	5.57±0.54	2.95±0.54
	Necrosis	0.21±0.07	0.00±0.00	8.01±1.92	90.05±2.26
		2 days
	Viable	97.59±0.63	97.25±0.58	81.84±0.98	1.47±0.66
	Apoptosis	2.41±0.63	2.75±0.58	4.84±0.73	3.52±1.53
	Necrosis	0.00±0.00	0.00±0.00	13.32±1.66	95.01±0.89
		3 days
	Viable	98.37±0.50	94.79±1.40	46.53±2.64	0.00±0.00
	Apoptosis	1.63±0.50	2.80±0.50	4.61±0.30	10.08±1.43
	Necrosis	0.00±0.00	2.41±0.70	48.86±2.43	89.92±1.43
		7 days
	Viable	98.00±0.57	76.52±1.02	0.00±0.00	0.00±0.00
	Apoptosis	1.88±0.55	7.59±0.53	9.44±0.91	10.10±0.15
	Necrosis	0.12±0.02	15.89±0.83	90.56±0.91	89.90±0.15

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[tfnI] Results presented as mean ± SD.