Platelet-concentrated and platelet poor-plasma promote different pattern on immunohistochemical expression of TGF-β1, however they impairs the osteoneogensis in calvarial defects treated with autograft due suppression of osteocalcin

Oliveira-Filho, Marco Antonio de; Souza, Marcelo; Tabushi, Fernando Issamu; Almeida, Luís Eduardo; Figueiredo, Fernanda Pirajá; Lourenço, Elora Sampaio; Giovanini, Allan Fernando; Malafaia, Osvaldo; Ribas Filho, Jurandir Marcondes

doi:10.1590/ACB360604

Marco Antonio de Oliveira-Filho

PhD. Medical Research Institute - Hospital Universitário Evangélico Mackenzie – Curitiba (PR), Brazil.

http://orcid.org/0000-0003-0016-4120

Marcelo Souza

MsC. Medical Research Institute - Hospital Universitário Evangélico Mackenzie – Curitiba (PR), Brazil.

http://orcid.org/0000-0002-1552-8399

Fernando Issamu Tabushi

PhD. Medical Research Institute - Hospital Universitário Evangélico Mackenzie – Curitiba (PR), Brazil.

http://orcid.org/0000-0002-3150-2164

Luís Eduardo Almeida

PhD. Assistant Professor. Marquette University - United States.

http://orcid.org/0000-0002-9802-3782

Fernanda Pirajá Figueiredo

Graduate student. Hospital Universitário Evangélico Mackenzie – Curitiba (PR), Brazil.

http://orcid.org/0000-0001-5982-7668

Elora Sampaio Lourenço

Graduate student. Hospital Universitário Evangélico Mackenzie – Curitiba (PR), Brazil.

http://orcid.org/0000-0002-3281-0433

Allan Fernando Giovanini ^**Corresponding author: afgiovanini@gmail.com | (55 41)3240-5488

PhD. Medical Research Institute - Hospital Universitário Evangélico Mackenzie – Curitiba (PR), Brazil.

http://orcid.org/0000-0002-1637-2955

Osvaldo Malafaia

PhD. Medical Research Institute - Hospital Universitário Evangélico Mackenzie – Curitiba (PR), Brazil.

http://orcid.org/0000-0002-1829-7071

Jurandir Marcondes Ribas Filho

PhD. Medical Research Institute - Hospital Universitário Evangélico Mackenzie – Curitiba (PR), Brazil.

http://orcid.org/0000-0002-5251-7672

*Corresponding author: afgiovanini@gmail.com | (55 41)3240-5488

Conflict of interest: Nothing to declare.

Author’s contribution

Substantive scientific and intellectual contributions to the study: Oliveira Filho MA, Tabushi FI, Souza M and Almeida LE; Conception and design: Oliveira Filho MA, Figueiredo FP, Lourenço ES and Souza M. Data acquisition, analysis and interpretation: Giovanini AF and Ribas Filho JM; Histopathological examinations: Giovanini AF; Statistics analysis: Oliveira Filho MA and Alemira LE; Manuscript preparation: Giovanini AF; Manuscript writing: Giovanini AF, Figueiredo FP and Lourenço ES; Critical revision: Malafaia O; Final approval: Malafaia O.

Figures (4)
Tables (3)

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Figure 1
Schematic model of platelet-poor plasma and platelet concentrate preparation.

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Figrue 2
Micrographs (a-e) demonstrate the histological characteristics in the ABG, ABG + PC, ABG + PPP, PPP and control groups, respectively, two weeks after surgery. A, B and C reveal similar presence of autograph in the ABG, ABG + PC and ABG + PPP groups, while connective tissue was predominant in the PPP (D) and control (E) groups. Micrographs (f-j) demonstrate the same groups four weeks after surgery. The larger bone deposition on the defect treated with only ABG is evident, while the osteoneogenesis was lower for the groups that received PC and PPP. The patterns of the PPP and control groups at four weeks were similar, but in the control group there was a scarce presence of bone matrix in the host bone. These patterns were similar to the sixth week (k-o). In this period, the intense presence of bone matrix in the ABG group may be seen. In the groups that received ABG + PC and ABG + PPP, the presence of osteogenesis was lower, while for the PPP and control groups the robust parts of the defects were composed of fibrotic tissue (hematoxylin and eosin stain, original magnification ×100).

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Figure 3
Micrographs (a-e) demonstrate the pattern of distribution of immunoexpression of TGF-ß1 (brownish color) in the ABG, ABG + PC, ABG + PPP, PPP and control groups, respectively, two weeks after surgery. Verify the intense presence of TGF-ß1 in ABG+PC group, while for the specimens in which PPP was inserted the immunopositivity for TGF-ß1 was scarce. Micrographs (f-j) demonstrate the same groups four weeks after surgery. The patterns of TGF-ß1 were suppressed in PPP groups, but the TGF-ß1 remains intense in specimens that received PC. These patterns were also similar to the sixth week (k-o). However, it was verified that the cytokine increased in extravascular area in specimens that received PPP. Micrographs (f-j) demonstrate the same groups on the sixth week after surgery. Verify the similar pattern of distribution of osteocalcin when compared to earlier stage of bone repair (second week) (original magnification ×100).

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Figure 4
Micrographs A, B, C, D and E demonstrate the pattern of distribution of immunoexpression of OC (brownish color) in the ABG, ABG + PC, ABG + PPP, PPP and control groups, respectively, two weeks after surgery. Verify the intense presence of OC on ABG+PC group while in the specimens in which PC or PPP was inserted the immunopositivity for OC was fewer (brownish color). However, it was evident that specimens that were treated with PC revealed higher OC when compared to specimens that received PPP, associated or not to ABG (original magnification ×250).

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Table 1
Histomorphometric analysis among the groups regarding bone matrix formation^* * Non-parametric Kruskal-Wallis test was employed; ,^** ** p < 0.05 indicates statistical significance; PPP: platelet-pour plasma; ABG: autogenous particulate bone graft;PC: platelet concentrate; PRP: platelet-rich plasma. .

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Table 2
Percentage of immunohistochemical presence of TGF-ß1 analysis among the groups^* * Non-parametric Kruskal-Wallis test was employed; ,^** ** p< 0.05 indicates statistical significance; PPP: platelet-pour plasma; ABG: autogenous particulate bone graft; PC: platelet concentrate .

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Table 3
Percentage of immunohistochemical presence of osteocalcin analysis among the groups*,^** ** p< 0.05 indicates statistical significance; PPP: platelet-pour plasma; ABG: autogenous particulate bone graft; PC: platelet concentrate .

Time period			PPP	PPP+Autograft	PRP+Autograft	Autograft	Control
2 Weeks			2.91 ± 0,75	5.61 ± 0,87	6.08 ± 1.03	6.61 ± 1.46	2.84 ± 0.29
	PPP	2.91 ± 0.75	----	< 0.001	< 0.001	< 0.001	0.916
	ABG+PPP	5.61 ± 0.87		----	= 0.461	= 0.123	< 0.001
	ABG+PC	6.08 ± 1.03			----	= 0.397	< 0.001
	ABG	6.61 ± 1,.46				----	< 0.001
	Control	2.84 ± 029					----
			5.01 + 0.93	6.08 ± 1.69	6.0 ± 1.4	8.24 ± 1.18	5.27 ± 1.02
4 Weeks	PPP	5.01 ± 0.93	----	= 0.207	= 0.242	< 0.001	= 0.743
	ABG+PPP	6.08 ±1.69		----	= 0.922	= 0.018	= 0.341
	ABG+PC	6.0 ± 1.4			----	= 0.015	= 0.390
	ABG	8.24 ± 1.18				----	= 0.002
	Control	5.27 ± 1.02					----
			7.52 ± 0.64	7.73 ± 1.41	7.53 ± 1.3	9.64 ± 1.43	7.86 ± 0.57
6 Weeks	PPP	7.52 ± 0.64	----	= 0.709	= 0.983	= 0.002	= 0.551
	ABG+PPP	7.73 ± 1.41		----	= 0,725	= 0.004	= 0.821
	ABG+PC	7.53 ± 1.3			----	= 0.002	= 0.565
	ABG	9.64 ± 1.43				----	= 0.006
	Control	7.86 ± 0.57					----

Time period			PPP	ABG+PPP	ABG + PC	ABG	Control
2 Weeks			12.65 ± 1.73	38.44 ± 2.61	76.05 ± 3.11	51.14 ± 2.38	17.22 ± 1.67
	PPP	12.65 ± 1.73	----	< 0.001	< 0.001	< 0.001	0.046
	ABG+PPP	38.44 ± 2.61		----	= 0.006	= 0.018	< 0.001
	ABG +PC	76.15 ± 3.11			----	= 0.048	< 0.001
	ABG	51.14 ± 2.38				----	< 0.001
	Control	17.22 ± 1.67					----
4 Weeks			3.84 ± 1.02	3.08 ± 1.69	76.08 ± 2.26	48.11 ± 1.74	45.27 ± 2.22
	PPP	3.84 ± 1.02	----	= 0.746	< 0.001	< 0.001	< 0.001
	ABG+PPP	3.08 ± 1.69		----	< 0.001	< 0.001	< 0.001
	ABG +PC	76.08 ± 2.26			----	= 0.015	= 0.028
	ABG	48.11 ± 1.74				----	= 0.704
	Control	45.27 ± 2.22					----
6 Weeks			7.42 ± 0.38	4.83 ± 1.66	69.82 ± 1.37	37.77 ± 2.17	48.21 ± 0.99
	PPP	7.42 ± 0.38	----	= 0.062	< 0.001	< 0.001	< 0.001
	ABG+PPP	4.83 ± 1.66		----	< 0.001	< 0.001	< 0.001
	ABG +PC	69.82 ± 1.37			----	= 0.024	= 0.003
	ABG	37.77 ± 2.17				----	= 0.066
	Control	48.21 ± 0.99					----

Time period			PPP	ABG+PPP	ABG + PC	ABG	Control
2 Weeks			8.32 ± 0.42	26.19 ± 2.89	34.07 ± 2.51	76.89 ± 3.05	44.62 ± 1.18
	PPP	8.32 ± 0.42	----	< 0.001	< 0.001	< 0.001	0.046
	ABG+PPP	26.19 ± 2.89		----	= 0.624	< 0.001	= 0.002
	ABG +PC	34.07 ± 2.51			----	< 0.018	= 0.026
	ABG	76.89 ± 3.05				----	= 0.002
	Control	44.62 ± 1.18					----
4 Weeks			71.16 ± 2.33	17.11 ± 1.49	31.84 ± 2.83	71.16 ± 2.33	43.11 ± 1.94
	PPP	4.38 ± 0.87	----	= 0.046	< 0.001	< 0.001	< 0.001
	ABG+PPP	17.11 ± 1.49		----	< 0.018	< 0.001	< 0.001
	ABG +PC	31.84 ± 2.83			----	= 0.022	= 0.028
	ABG	71.16 ± 2.33				----	= 0.044
	Control	43.11 ± 1.94					----
6 Weeks			3.82 ± 0.19	14.28 ± 1.42	27.28 ± 1.66	69.08 ± 2.73	42.21 ± 0.99
	PPP	3.82 ± 0.19	----	= 0.006	< 0.001	< 0.001	< 0.001
	ABG+PPP	14.28 ± 1.42		----	= 0.018	< 0.001	< 0.001
	ABG +PC	27.28 ± 1.66			----	= 0.002	= 0.003
	ABG	69.08 ± 2.73				----	= 0.044
	Control	42.21 ± 0.99					----

[1] *Corresponding author: afgiovanini@gmail.com | (55 41)3240-5488

Brasil

Brasil

Platelet-concentrated and platelet poor-plasma promote different pattern on immunohistochemical expression of TGF-β1, however they impairs the osteoneogensis in calvarial defects treated with autograft due suppression of osteocalcin

ABSTRACT

Purpose

Methods

Results

Conclusions