Impact of Hyaluronic Acid on the Viability of Mesenchymal Cells Derived from Adipose Tissue Grown in Collagen Type I/III Membrane

Kaleka, Camila Cohen; Debieux, Pedro; Antonioli, Eliane; Zucconi, Eder; Cohen, Moisés; Ferretti, Mário

doi:10.1055/s-0041-1740198

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Original Article • Rev. bras. ortop. 57 (6) • Nov-Dec 2022 • https://doi.org/10.1055/s-0041-1740198 copy

Impact of Hyaluronic Acid on the Viability of Mesenchymal Cells Derived from Adipose Tissue Grown in Collagen Type I/III Membrane

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

Objective

To evaluate in vitro the viability of mesenchymal stem cells derived from adipose tissue (AD-MSCs) in different commercial solutions of hyaluronic acid (HA) before and after being sowed in collagen I/III membrane.

Methods

In the first stage, the interaction between AD-MSCs was analyzed with seven different commercial products of HA, phosphate buffered saline (PBS), and bovine fetal serum (BFS), performed by counting living and dead cells after 24, 48 and 72 hours. Five products with a higher number of living cells were selected and the interaction between HA with AD-MSCs and type I/III collagen membrane was evaluated by counting living and dead cells in the same time interval (24, 48 and 72 hours).

Results

In both situations analyzed (HA + AD-MSCs and HA + AD-MSCs + membrane), BFS presented the highest percentage of living cells after 24, 48 and 72 hours, a result higher than that of HA.

Conclusion

The association of HA with AD-MSCs, with or without membrane, showed no superiority in cell viability when compared with BFS.

Keywords
cartilage; articular; mesenchymal stem cells; mesenchymal stem cell transplantation; hyaluronic acid

Number	Product	Molecule	Concentration	Molecular weight (MDa)	Elasticity (Pa 2.5 hz)	Viscosity (Pa 2.5 hz)	Source	Crosslink molecule
1	Opus Joint	Sodium hyaluronate	1.5%	5.4	145	100	Bacterial	No
2	SupraHyal Duo	Sodium hyaluronate	1%	0.75	1.2		Bacterial	No
3	Fermathron	Sodium hyaluronate	1%	1.19–2.03	0.84		Bacterial	No
4	Orthovisc	Sodium hyaluronate	1.5%	1–2.9	60	46	Bacterial	No
5	Synolis VA	Sodium Hyaluronate + sorbitol	2% + 4%	2	329	143	Bacterial	No
6	Synvisc	Hilano GF-20	0.8%	6	111	25	Avian	yes
7	Osteonil	Sodium hyaluronate	1%	1–2	−	210	Bacterial	No

Moment	Treatment	Average % of living cells		SD of % of living cells
Moment	Treatment	Mean (95%CI)	p-value	Mean (95%CI)	p-value
24h	HA 1	93.7 (90.3–97.3)	0.075	2.7 (1.4–5.3)	0.956
	HA 2	94.4 (91.5–97.5)	0.169	2.9 (1.4–6.0)	0.839
	HA 3	93.4 (91.4–95.4)	0.090	2.1 (1.0–4.2)	0.707
	HA 4	92.9 (88.4–97.6)	< 0.001	3.7 (2.1–6.5)	0.470
	HA 5	96.2 (93.3–99.3)	0.704	1.7 (0.8–3.5)	0.669
	HA 6	95.3 (91.0–99.8)	0.084	2.2 (1.0–5.1)	0.854
	HA 7	94.7 (92.2–97.4)	0.041	2.5 (1.1–5.6)	0.908
	Middle	88.2 (84.1–92.5)	< 0.001	3.8 (2.2–6.5)	0.276
	PBS	97.3 (93.2–100.0)	Reference	2.6 (0.6–11.4)	Reference
48h	HA 1	95.7 (91.7–99.9)	0.097	2.4 (0.7–7.5)	0.095
	HA 2	94.5 (91.2–97.8)	0.001	1.2 (0.8–1.9)	0.865
	HA 3	94.1 (91.6–96.6)	< 0.001	2.6 (1.7–4.0)	0.272
	HA 4	95.0 (90.6–99.7)	0.059	3.0 (1.6–5.5)	< 0.001
	HA 5	95.8 (93.4–98.3)	0.089	2.8 (1.5–5.3)	< 0.001
	HA 6	93.5 (88.7–98.5)	0.003	3.4 (1.7–6.7)	< 0.001
	HA 7	92.7 (87.1–98.6)	0.011	2.3 (1.8–2.9)	0.173
	Middle	86.1 (81.9–90.4)	< 0.001	5.7 (2.8–11.5)	0.002
	PBS	98.1 (96.2–100.0)	Reference	1.3 (0.4–4.6)	Reference
72h	HA 1	93.9 (88.5–99.8)	0.049	3.2 (1.3–8.2)	0.105
	HA 2	94.3 (91.8–96.8)	< 0.001	3.7 (2.4–5.8)	0.308
	HA 3	93.6 (91.7–95.4)	< 0.001	5.6 (2.6–12.4)	0.123
	HA 4	91.6 (86.7–96.8)	0.002	3.1 (1.5–6.3)	0.299
	HA 5	88.5 (80.3–97.5)	0.007	7.3 (3.4–15.6)	0.025
	HA 6	93.3 (89.4–97.3)	0.002	4.6 (2.5–8.4)	0.059
	HA 7	92.7 (90.0–95.4)	< 0.001	2.9 (2.3–3.7)	0.534
	Middle	82.6 (70.3–97.2)	0.009	4.4 (2.7–7.2)	0.091
	PBS	98.6 (97.2–100.0)	Reference	2.1 (0.6–7.4)	Reference

Moment	Treatment	Estimated mean (95%CI)	p-value
24h	BFS	95.8 (93.4–97.4)	< 0.001
	Acid 7	86.3 (77.7–91.5)	0.826
	Acid 5	81.3 (72.3–87.3)	0.350
	Acid 4	86.6 (81.8–90.2)	0.801
	Acid 2	81.2 (61.8–90.7)	0.429
	Acid 1	83.7 (74.4–89.6)	0.555
	PBS	87.8 (74.5–94.1)	Reference
	BFS	95.0 (93.3–96.3)	0.395
	Acid 7	93.0 (88.9–95.5)	0.755
	Acid 5	86.9 (79.3–91.8)	0.017
48h	Acid 4	88.6 (78.6–93.9)	0.222
	Acid 2	92.9 (89.8–95.0)	0.699
	Acid 1	89.5 (86.7–91.7)	0.033
	PBS	93.8 (90.9–95.7)	Reference
	BFS	95.6 (93.3–97.0)	0.063
	Acid 7	89.7 (84.8–93.0)	0.765
	Acid 5	86.1 (78.2–91.1)	0.749
72h	Acid 4	90.8 (81.7–95.4)	0.674
	Acid 2	92.5 (87.7–95.4)	0.298
	Acid1	85.5 (79.7–89.6)	0.671
	PBS	88.4 (75.9–94.4)	Reference

Moment	Comparison			Corrected p-value
	BFS	X	Acid 7	0.105
	BFS	X	Acid 5	0.003
	BFS	X	Acid 4	< 0.001
	BFS	X	Acid 2	0.497
	BFS	X	Acid1	0.028
	Acid 7	X	Acid 5	0.497
24h	Acid 7	X	Acid 4	>0.99
	Acid 7	X	Acid 2	>0.99
	Acid 7	X	Acid1	>0.99
	Acid 5	X	Acid 4	0.497
	Acid 5	X	Acid 2	>0.99
	Acid 5	X	Acid1	>0.99
	Acid 4	X	Acid 2	>0.99
	Acid 4	X	Acid1	>0.99
	Acid 2	X	Acid1	>0.99
48h	BFS	X	Acid 7	>0.99
	BFS	X	Acid 5	0.017
	BFS	X	Acid 4	>0.99
	BFS	X	Acid 2	>0.99
	BFS	X	Acid 1	0.033
	Acid 7	X	Acid 5	0.197
	Acid 7	X	Acid 4	>0.99
	Acid 7	X	Acid 2	>0.99
	Acid 7	X	Acid 1	>0.99
	Acid 5	X	Acid 4	>0.99
	Acid 5	X	Acid 2	>0.99
	Acid 5	X	Acid 1	>0.99
	Acid 4	X	Acid 2	>0.99
	Acid 4	X	Acid 1	>0.99
	Acid 2	X	Acid 1	0.204
	BFS	X	Acid 7	0.002
72h	BFS	X	Acid 5	0.012
	BFS	X	Acid 4	>0.99
	BFS	X	Acid 2	>0.99
	BFS	X	Acid1	<0.001
	Acid 7	X	Acid 5	>0.99
	Acid 7	X	Acid 4	>0.99
	Acid 7	X	Acid 2	>0.99
	Acid 7	X	Acid1	>0.99
	Acid 5	X	Acid 4	>0.99
	Acid 5	X	Acid 2	>0.99
	Acid 5	X	Acid1	>0.99
	Acid 4	X	Acid 2	>0.99
	Acid 4	X	Acid1	>0.99
	Acid 2	X	Acid1	0.213

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[1] Address for correspondence Camila Cohen Kaleka, MD, PhD, Av. Albert Einstein, 627/701, Bloco A1–sala 306, São Paulo, SP, 05652-900, Brasil, camilakaleka@gmail.com